Ring a unsaturated 6, 16-dimethyl steroids of the pregnane series



United States Patent Ofiiice 3,054,811 Patented Sept. 18, 1962 3,054,811 RING A UNSATURATED 6,16-DHMETHYL STER- ID 0F TIE PREGNANE ERIES Glen E. Arth, Cranford, N.J., Roger E. Beyler, Carbondale, 11L, and Lewis H. Sarett, Princeton, N.J., assignors to Merck & Co., Inc., Railway, Ni, a corporation of New Jersey No Drawing. Filed Sept. 14, 1961, Ser. No. 138,007 2 Claims. (Cl. 260397.45)

This invention is concerned generally with novel steroid compounds and with processes of preparing the same. More particularly, it relates to novel ring A unsaturated 1700-hydroxy-11-oxygenated-2O-keto-steroids of the pregnane series having methyl substituents attached to both the C-6 and C-l6 carbon atoms, as for example 6,16-

dimethyl 11 oxygenated 1,4 pregnadiene 1700,21- diol 3,20 dione compounds, 6,16-dimethyl-11-oxygenated-4-pregnene-17u,21-diol-3,20-dione compounds, and to the process of preparing these .compounds starting with 16 methyl 4 pregnene 1700,21 diol 3,11,20- trione. These novel ring A unsaturated 6,16-dimethyl- 170z-hydroxy-11-oxygenated-20-keto-steroids of the pregnane series possess extremely high anti-inflammatory activity, and are especially effective for the treatment of arthritis and related diseases since they can be administered for their cortisone-like action in extremely low dosage thereby minimizing undesired side effects.

This is a continuation-in-part of application Serial No. 683,923, new Patent No. 3,004,994, filed September 16, 1957.

The novel 6,16-dimethyl-1l-oxygenated-l,4-pregnadiene-17 x,21-diol-3,20-dione compounds, included in the present invention, may be chemically represented as follows:

CHzOR wherein X is hydrogen or halogen, R stands for hydrogen or acyl, and Z is a keto or hydroxy substituent.

These 6,16 dimethyl-1l-oxygenated-1,4-pregnadiene- 1700,21-diol-3,20-dione compounds are prepared by reacting 16 methyl 4 pregnene 1700,21 diol 3,11,20- trione, more particularly 1600-methy1-4-pregnene-1700,2ldiol 3,11,20 trione; 16,8 methyl 4 pregnene 1700, 21-diol-3,11,20-trione; with formaldehyde under acidic conditions to form 1700,20,20,21-bismethylenedioxy-16- methyl-4-pregnene-3,1l-dione, more particularly 1700,20, 20,21-bismethylenedioxy 1600 methyl 4 pregnene- 3,11 dione; 1700,20,20,21 bismethylenedioxy 16B- methyl-4-pregnene-3,ll-dione; which is reacted with ethylene glycol in the presence of an acidic catalyst to produce 3 ethylenedioxy 1700,20,20,21 bismethylenedioxy-16-methyl-5-pregnene-ll-one, more particularly 3- ethylenedioxy 1700,20,20,21 bismethylenedioxy 1600- methyl 5 pregnene 11 one; 3 ethylenedioxy 1700, 20,20,21 bismethylenedioxy 165 methyl 5 pregnene-ll-one. The latter compound is reacted with perbenzoic acid, perphthalic acid and the like, thereby forming 3 ethylenedioxy l700,20,20,21 bismethylenedioxy- 5,6 epoxy 16 methyl pregnane 11 one, more particularly 3 ethylenedioxy 1700,20,20,21 bismethylenedioxy 5,6 epoxy 1600 methyl pregnane 11 one; 3 ethylenedioxy 1700,20,20,21 bismethylenedioxy 5,6- epoxy-l6B-methyl-pregnane-1l-one which is then reacted with formic acid to produce a mixture of 1700,20,20,21- bismethylenedioxy 16 methyl 6 formyloxy pregnane-5-ol-3,1l-dione, more particularly 1700,20,20,21 bismethylenedioxy 1600 methyl 6 formyloxy pregnane 5 ol 3,11 dione; 1700,20,20,21 bismethylenedioxy 16,8 methyl 6 formyloxypregnane 5 o1- 3,11-dione; and 1700,20,20,21 bismethylenedioxy 16- methyl 5 formyloxy pregnane 6 ol 3,11 dione, more particularly 1700,20,20,21-bismethylenedioXy-1600 methyl 5 formyloxy pregnane 6 ol 3,11 dione; 1700,20,20,21 bismethylenedioxy 16,8 methyl 5- forrnyloxy-pregnane-6-ol-3,1l-dione, which mixture, upon reaction with an aqueous alkali hydroxide solution, is converted to 1700,20,20,21-bismethylenedioxy-l6-methy1- allopregnane-3,6,1l-trione, more particularly, 1700,20,20, 21 bismethylenedioxy 1600 methyl allopregnane- 3,6,11 trione; 1700,20,20,21 bismethylenedioxy methyl-allopregnane3,6,ll-trione. The last-named compound is reacted with butanone dioxolane to produce 3 ethylenedioxy 1700,20,20,21 bismethylenedioxy 16- methyl-allopregnane-6,ll-dione, more particularly 3- ethylenedioxy 1700,20,20,21 bismethylenedioxy 1600- methyl allopregnane 6,11 dione; 3 ethylenedioxy- 1700,20,20,21 bismethylenedioxy 166 methyl allopregnane-6,11-dione which is reacted with a methyl Grignard reagent to form 3-ethylenedioxy-l700,2O,20,21-bismethylenedioxy 600,16 dimethyl allopregnane 6B olll-one, more particularly 3-ethylenedioxy-1700,20,20,21- bismethylenedioxy 600,1600 dimethyl allopregnane 6B- 01 11 one; 3 ethylenedioxy 1700,20,20,21 bismethylenedioxy 600,165 dimethyl allopregnane 65 ol 11- one. This 3ethylenedioxy-1'700,20,20,21-bismethylenedioxy 600,16 dimethyl allopregnane 68 01 11 one, more particularly 3-ethylenedioxy-1700,20,20,21-bisrnethylenedioxy 600,1600 dimethyl allopregnane 613 01 11- one; 3 ethylenedioxy 1700,20,20,21 bismethylenedioxy-600,16B-dimethyl-allopregnane-dfi-ol-1l-one is reacted With a dehydrating agent such as thionyl chloride in pyridine to form the corresponding 3-ethylenedioxy- 1700,20,20,21 bismethylenedioxy 6,16 dimethyl 5- pregnene-ll-one, more particularly 3-ethylenedioxy-1700, 20,20,21 bismethylenedioxy 6,1600 dimethyl 5- pregnene 11 one; 3 ethylenedioxy 1700,20,20,21 bismethylenedioxy 6,165 dimethyl 5 pregnene 11- one, which, upon reaction with p-toluenesulfonic acid monohydrate in acetone is converted to l700,20,20,21- bismethylenedioxy 600,16 dimethyl 4 pregnene 3,11- dione, more particularly 1700,20,20,2l-bismethylenedioxy- 600,1600 dimethyl 4 pregnene 3,11 dione; 1700,20, 20,21 bismethylenedioxy 600,165 dimethyl 4 pregnene-3,11-dione; the latter compound is reacted with an aqueous organic acid hydrolyzing agent to form 600,16- dimethyl 4 pregnene 1700,21 diol 3,11,20 trione, more particularly 600,1600 dimethyl 4 pregnene 1700, 21 diol 3,11,20 trione; 600,16fi-dimethyl 4 pregnene- 1700,21-diol-3,11,20-trione which can be reacted with an acylating agent to form the corresponding 21-acylate. Alternatively, the 3-ethylenedioxy 1700,20,20,21 bismethylenedioxy 6,16 dimethyl 5 pregnene 11 one, more particularly 3-ethylenedioxy-1700,20,20,21 bismethylenedioxy 6,1600 dimethyl 5 pregnene 11- one; 3 ethylenedioxy 17 00,20,20,21 bismethylenedioxy- 6,16fi-dimethyl-5-pregnene-1l-one, above-mentioned, can be reacted with lithium aluminum hydride thereby reducing the ll-keto substituent to an lie-hydroxy radical to produce 3-ethylenedioxy-1700,20,20,21-bismethylenedioxy 6,16 dimethyl 5 pregnene 11p 01, more particularly 3 ethylenedioxy 1700,20,20,21 bismethylenedioxy 6,160: dimethyl 5 pregnene 11,8 ol; 3 ethylenedioxy 17a,20,20,21 bisrnethylenedioxy 6,1613 dimethyl-S-pregnene-llB-ol which is converted, by reaction with p-toluenesulfonic acid monohydrate in acetone, to 17a,20,20,21blSII16thylCI16-dl0XY-6a,16 dimethyl 4- pregnene-1lfi-ol-3-one, more particularly 17oc,20,20,21- bismethylenedioxy 60:,16a dirnethyl 4 pregnene 11,6- ol 3 one; 17a,20,20,21 bismethylenedioxy 6a,16;8- dimethyl-4-pregnene-11p-ol-3-one; the latter compound is reacted With an aqueous organic acid hydrolyzing agent thereby forming 60:,16-dimethyl-4-pregnene-11,3,17a,21- triol-3,20-dione, more particularly 60:,160t-dit1'lfithYl-4- pregnene-llp,l7a,2l-triol-3,20-dione; 60;,165 dimethyl- 4-pregnene-11p,17 x,21-triol-3,20-dione which can be reacted with an acylating agent thereby forming the corresponding 2l-acylate derivative.

I Alternatively, the 3-ethylenedioxy 17a,20,20,21 bismethylenedioXy-5,6-epoxy-l6-rnethyl pregnane ll one, more particularly 3-ethylenedioxy-17a,20,20,21-bismethylenedioxy-5,6-epoxy-16u-methyl-pregnane-1l-one; 3-ethylenedioxy-17a,20,20,2l-bismethylenedioxy-S,6-epoxy 165- methyl-pregnane-lhoue is reacted With methyl magnesiurn bromide to form 17u,20,20-2l-bisrnethylenedioxy- 6,8,16-dimethyl-3ethylenedioxy-allopregnane-5a o1 11- one; more particularly 17a,20,20,2l-bisrnethylenedioxy- 65,16a-dirnethyl-3-ethylenedioXy-allopregnane-Sa-OI 11- one; 17a,20,20,21-bismethylenedioxy-6B,16/3-dimethyl-3- ethylenedioxy-allopregnanefioc-ol 11 one; which, upon reaction with adilute aqueous methanolic sulfuric acid solution is converted to 17a,20,20,2l-bismethylenedioxy- 6,8,16-din1ethyl-allopregnane-3,1l-dione-Sa-ol; more particularly 17a,20,20,21-bismethylenedioXy-6fi,l6a-dimethyl-allopregnane-3 ,1 l-dione-Sa-ol; l7ot,20,20,2l-bismethy1- enedioxy-6fi,lfifi-dixnethyl-allopregnane-B,1l-dione-Sa 01. The last-named compound, 'When reacted with dilute methanolic KOH solutionteg. 0.25%) under reflux for approximately one-half hour, is converted to l7oc,20,2(), 2l-bismethylenedioXy-6fi,l6-dirnethyl-4-pregnene-3,11 dione; more particularly l'7a,20,20,2l-bismethylenedioxy- 6,3, l6ot-dimethyl-4pregnene-3,1 l-dione; 17a,20,20,2l-bisinethylenedioxy-6fl,l6p-dimethyl-4-pregnene-3,l1 dione; the latter compound is reacted with an aqueous organic acid hydrolyzing agent (in accordance with the" procedures set forth hereinabove in connection with the treatment of 17a,20,20,2l-bismetl'1ylenedioxy-6a,16-dimethyl- 4-pregnene-3,11-dione) to form 6,8,16-dimethyl-4-pregnene-17a,21-diol-3,11,20-trione; more particularly 65, 16a-dimethyl-4-pregnene-17 1,21-diol 3,l1,20 trione; 6,8, 16fi-dimethyl-4-pregnene-17ot,2l-diol-3 ,11,20-trione; which can be reacted with an acylating agent such as acetic anhydride to form the corresponding 2l-acylate, more particularly the 21-acetate. Instead of hydrolyziug' the 17cz,20,20,2 l-bismethylcnedioxy-6fl,16-dimethyl 3 ethylenedioxy allopregnane II-OIlB-Soz-Ol by treatment with dilute aqueous sulfuric acid, the above-named compound can be reacted with lithium aluminum hydride thereby reducing the ll-keto substituent to an llfi-hydroxy radical to produce 17m,20,20,21-bismethylenedioXy-6fl,16-dirnethyl-3-ethylenedioxy allopregnane-S a,11 3 diol, more particularly 17a,20,20,2l-bismethylenedioxy 6 3,160: dimethyl-3ethylentedioxy-allopregnane-5a,1 1,6-diol; 17 :,20, 20,21-bismethylenedioXy-6B,16fl-dimethyl-3-ethylene dioxyallopregnane-a,1lfi-diol; which is converted by reaction with dilute aqueous methanolic sulfuric acid solution to 17a,20,20,21-bismethylenedioxy-6fl,16 dimethylallopregnane-5a,1lfl-diol-S-one, more particularly 170;,20, 20,21-bismethylenedioxy-6,8,la-dimethyl allopregnane- 5a,11/3-diol-3-one; 17a,20,20,21 bismethylenedioxy 6ft, 16fl-dimethyl-aIlQpregnane-Sa,1lfl-diol-3-one. The latter compound, when reacted with dilute methanolic potassiumhydroxide solution (e.g. 025%) under reflux for about one-half hour, is converted to 17a,2Q,20,Z1-bi$ methylenedioxy-6B,16-dimethyl-4-pregnened16-01-3 one, more particularly 17a,20,20,21-bismethylenedioxy-6B,16adimethyl-4-pregnene-l1,3-ol-3 one; 17,20,20,2 1-bismeth ylenedioxy-GB,165-dirnethyl-4-pregnene-1 1 S-ol-3-one; the latter compound is reacted with an aqueous organic acid hydrolyzing agent thereby forming 65,16-dimethyl-4- pregnene-ll,8,17u,21-triol-3,20 dione, more particularly 6,8,16u-dimethyl-4-pregnene-11,6,17a,21-triol-3,20 dione; 6B,16fi-dimethyl-4-pregnene-11;8,17u,21-triol-3,20 dione; which can be reacted with an acylating agent such as acetic anhydride thereby forming the corresponding 21- acylate derivative such as 6(3,l6u-dimethyl-4-pregnene- 115,l7ct,2l-triol-3,20-dione 21-acetate; 6,3,16,6-dimethyl- 4-pregnene-1 1,8, 1 7a,21-triol-3,20-dione 21-acetate.

This 6,16-dirnethyl-4-pregnene-1 1 13,17oc,21-t1i0l3,20-di one ZI-acylate, more particularly 6a,16u-dimethyl-4-preg- 116I1811,8,l70c,21-t1'i0l 3,20 dione 2l-acylate; 60:,16fi-dimethyl-4-pregnene-1113,17a,21-triol-3,20-dione 21-acylatc; 6,6,l6a-dimethyl-4-pregnene-11fi,17a,21-triol-3,20 dione 21-acylate; 6,3,16fl-dimethyl-4-pregnene-11;8,17 x,21-triol- 3,20-dione 21-acylate; is reacted with a dehydrating agent such as methane sulfonyl chloride in pyridine, or phosphorous oxychloride dissolved in pyridine, to produce the corresponding 6,16-dimethyl-4,9(11)-pregnadiene-17u, 21-dio1-3,20-d ione 21-acy1ate, more particulary 60:,1611- dimethyl-4,9(1l)-pregnadiene-l7a,21-dio1-3,20dione 21- acylate; 6a,l6,8-dimethyl-4,9(l1)-p1'egnadiene-17a,21-diol-3,20-dione 21 acylate; 6fi,l6a-dimethyl-4,9(l1)-pregnadiene-l7a,21-diol-3,20-dione 2l-acylate; 6;8,16fl-dimethyl-4,9( 11 )-pregnadiene-17et,21-dio1-3,20-dione 21-acy1ate; the latter compound is reacted with hypobromous acid (cg. N-bromosuccinirnide and perchloric acid) to produce 9a-brorno-6, 1 6-dimethyl-4-pregnene-1 1fi,17m,21-t1'i01- 3,20-dione 21-acylate, more particularly 9cz-b1'0m0-6oc, 16a-dimethyl-4-pregnene-1lfi,17a,2l-triol-3,20 dione 21- acylate; 9a-bromo-6a,16B-dimethyl-4-pregnene 1119,1701, 21-triol-3,20-dioue 21-acylate; 9ot-bromo-65J6a-dimethyl- 4-pregnene-11;8,17a,21 triol 3,20 dione 21-acylate; 6,8, I6fi-dimethyl-4-pregnene-l1B,17a,21-triol-3,20 dione 2lacylate; which is reacted with anhydrous potassium acetate in ethanol to produce 6,16-dimethyl-9,1l-epoxy-4- pregnene-l7m,21-diol-3,20 dione 21 acylate, more particularly 6a,16a-dimethyl-9,11-epoXy-4-pregnene ;,21- diol-3,20-dione 21-acylate; 6a,16fl-dimethy1-9,11- epoxy- 4-pregnene-l7a,21-diol-3,20-dione 2l-acylate; 66,16u-dimethyl-9,11-epoxy-4-pregnene-17a,21-diol-3,ZO-dione 21- acylate; 6/3,16,8-dirnethyl-9,1l-epoxy-4-pregnene 1701,21- di0l-3,20-dione 21-acylate. This 9,1l-epoxide is then reacted with hydrogen fluoride in tetrahydrofuran to produce 6,16-dimethyl-19a-fiuoro-4-pregnene 11,6,17 ce,21- triol-3,20-dione 21 acylate, more particularly 6a,16ccdimethyl-9 ot-fiuoro-4-pregnene-1 1p, 17a,21-triol-3,20-dione 21-acylate; 6a,16,8-dimethyl-9ufiuoro 4 pregnene 11/3, 17a,21-triol-3,20-dione ZI-acylate; 6B,16u-dimethyl 9o:- fluoro-4-pregnene-11 6,17a,21-tri0l-3,20 dione 21-acy1ate; 6 8,16/3-dimethyl-9a-fiuoro-4-pregnene 11fi,17a,21 triol- 3,20-dione 21-acylate; this compound is reacted with a hydrolyzing agent to form 6,16-dimethyl-9a-fiuoro-4- pregnene-l113,17a,21-triol-3,20 dione, more particularly 6a,16u-dimethyl-9a-fluoro-4-pregnene 11;3,l7a,21 triol- 3,20-dione; 6a,16,8 dimethyl-9a-fiuoro-4 pregnene 115, 17a,21-triol-3,20-dione; 6p,16m-dimethyl-9oc-fluoro-4-pregnene-llfi,17a,21-trio1-3,20-dione; 6,8,16,8-dirnethyl-9a-fiuoro-4-pregnene-l1,8,17u,21-triol-3,20-dione; which can be reacted With an acylating agent to produce 6,16-dimethyl- 9a-fiuoro-4-pregnene-l 1B, l7 c,21-triol-3,20-dione 21-acylate, more particularly 6a,16m-dimethyl-9a-fluoro-4-pregnone-l 1[3,17a,2lt1i0l-3,20 dione 21 acylate; 612,16B-dimethyl-9a-fluoro-4-pregnene-11,3,17a,21-tri0l-3,20 dione Zl-acylate; 65,160t-dil'n6thYI-9a-flfl0l0 4 pregnene 11B, 17a,21-triol-3,20-dione 21-acylate; 65,165 dimethyl 9afluoro-4-pregnene-1 1 8,17a,21-triol-3,20-dione 21-acy1atc. This 6,16-dimethyl-9a-fluoro-4 pregnene 11;8,17a,21- triol-3,20-dione 2l-acylate, more particularly 6a,16a-dimethyl-9a-fluoro-4-pregnene-l1fi,17a,21-triol-3,20 dione 2l-acylate; 6a,16,8-dimethyl-9a-iluoro-4 pregnene 1148, 17a,21-triol-3,20-dione 21-acylate; 65,160 dimethyl 9:1 fiuoro-4-pregnene-11,5,17q,21-triol-3,20-dione 21-acylate;

618,16 8-dimethyl-9a-fluoro-4-pregnene 1lfi,17oc,21 triol- 3,20-dione 21-acylate; is reacted with chromium trioxide in pyridine to form 6,16-dimethyl-9a-fluoro-4-pregnene- 1706,21-diO1-3,11,20-t1'i01'le 2l-acylate, more particularly 60c, l6a-dimethyl-9a-fluoro-4-pregnene-17a,21-diol-3 ,1 1,20- trione; 6a,l6,8dimethyl-9a-tluoro-4-pregnene-17a,21-diol- 3,11,20-trione; 6,8,16u-dimethyl-9a-fluoro-4-pregnene-17a, 2l-diol-3,1 1,20-trione; 6,8,16,6-dimethyl-9u fiuoro-4-pregnene-17a,21-diol 3,11,20 trione; which, upon reaction with a hydrolyzing agent, forms 6,16-dimethyl-9a-fluoro- 4-pregnene-17a,2l-diol-3,11,20-trione, more particularly 6a, 160c-diI116thY1-9 a-fiuoro4-pregnenel7oc,2 1-diol-3 ,1 1,20- trione; 6a,l6fi-dimethyl-9oc-fluoro-4-pregnene-l7a,2l-diol- 3,11,20-trione; 65,16a-dimethyl-9u-fluoro-4-pregnene-17a, 21-diol-3,11,20-tri0ne; 6,8,16,8-dimethyl-9a-fluoro-4 pregnene-l7a,21-diol-3, l 1,20-trione.

The 6,16-dimethyl-11-oxygenated-4 pregnene 17a,21- diol-3,20-dione compounds prepared hereinabove (e.g. the 6,16-dimethyl-4-pregnene-17a,2l-diol-3,11,20 trione compounds and the 6,l6-dimethyl-4-pregnene-11p,17a,21- triol-3,20-dione compounds) are contacted with the dehydrogenating activity of Schizomycetes micro-organisms, or with selenium dioxide, thereby forming the corresponding 6,16-dimethyl-11-oxygenated-1,4 pregnadiene- 17u,21-dio1-3,20-dione compound, more particularly 60c, l6u-dimethyl-l,4pregnadiene-17a,2l-diol-3,11,20 trione; 6a,16m-dimethyl-1,4-pregnadiene-1 1B,17u,21-triol-3,20-dione; 6a,16a dimethyl-9a-fluoro-l,4-pregnadiene-17a,21- diol-3 ,11,20-trione; 6oz, 16a-dimethy1-9u-fluoro-1,4-pregnadiene 11,8,17a,21 triol-3,20-dione; 6u,16,8-dimethyl-1,4- pregnadiene-l7a,2l-diol-3,11,20-trione; 6u,16{5 dimethyl- 1,4-pregnadiene-ll,8,17a,21-triol 3,20 dione; 6a,16fidi methyl-9a-fluoro-1,4-pregnadiene-17a,21-diol-3,11,20 trione; 6a,l6B-dimethy1-9u-fluoro1,4-pregnadiene1 113,17, 21-triol-3,20-dione; 6,6,16a-dimethyl-1,4-pregnadiene-17a, 2l-diol-3,ll,20-trione; 6,8,l6a dimethyl-1,4-pregnadiene- 11 3,17u,2l-triol-3,2O-dione; 6B,l6a-dimethyl 90c fluoro- 1,4-pregnadiene-17a,2l-diol 3,11,20 trione; 6l3,l6a-di methyl-9a-fluoro-l,4-pregnadiene-l15,17a,21-trio1 3,20- dione; 6,8,16,8-dimethyl-l,4-pregnadiene-17a,21-diol-3,1l, 20-trione; 65,16B-dimethyl-L4 pregnadiene 11B,17 x,21- tIiol-3,20-dione; 66,166 dimethyl-9a-fluoro-1,4-pregnadiene-17a,21-diol-3,11,20-trione; 6 8, 1 65-dimethyl-9 OL-fllJOI'O- 1,4-pregnadiene-l 1 8, l7a,2 1-triol-3,20-dione.

The 16 methyl 4 preguene 170 21 diol 3,11,20- trione compounds used as starting materials in the abovementioned procedures are conveniently prepared starting with the known 16-pregnene-3a-ol-11,20-dione 3-acetate in accordance with the following procedure: 16-pregnene- 3a-ol-11,20-dione 3-acetate is reacted with methyl mag nesium iodide in the presence of cuprous chloride thereby forming l6a-methyl-pregnane-3a-ol-l1,20-dione 3-acetate, which is reacted with aqueous methanolic hydrochloric acid to form l6a-methyl-pregnane-3a-ol-l1,20-dione. The latter compound, which is a potent anaesthetic, is reacted with acetic anhydride in the presence of p-toluene-sulfonic acid catalyst to form a mixture of enol acetates containing 16a-methyl-l7(20)-pregnene-3a,20-diol-1l-one 3,20- diacetate; this mixture, after chromatographic purification to remove any unchanged starting material, is reacted with perbenzoic acid and the resulting l6u-methyl-17a,20- epoxy-pregnane-3a,20-diol-1l-one 3,20-diacetate is hyrolyzed With methanolic potassium bicarbonate to produce l6u-methyl-pregnane-3a,17a-diol-11,20-dione. The latter compound is reacted with bromine in chloroform to form 2l-bromo-16u-methyl-pregnane-3a,17u-diol-11, 20-dione which is reacted with sodium iodide in acetone to produce 21-iodo-16a-methyl-pregnane-3a,l7a-diol-ll, 20-dione which is converted without isolation to l6ot-methyl-pregnane-3a,17u,21-triol-11,20-dione 2l-acetate by reaction with anhydrous potassium acetate; this compound is reacted with chromium trioxide in pyridine to form 1600 methyl pregnane 17a,21 diol 3,11,20 trione Zl-acetate. The 16a-methyl-pregnane-l7a,21-dio1-3,11, 20-trione 21-acetate is reacted with bromine in glacial acetic acid-chloroform to produce 4-bromo-16a-methyl pregnane-17a,21-diol-3,l1,20-trione, which is then reacted with semicarbazide to form 16a-methyl-4-pregnene-17a, 2l-diol-3,ll,20-trione 3,20-bis-semicarbazone 21-acetate. T he l6a-methyl-4-pregnene-17u,2t1-di0l-3,11,20-trione-3, 20-bis-semicarbazone 21-acetate is reacted with potassium bicarbonate or potassium hydroxide in aqueous. methanol to form 16u-methyl-4-pregnene-17a,21-diol-3,11,20-trione 3,20-bis-semicarbazone which is then hydrolyzed under acid conditions toproduce 16a-methyl-4-pregnene-1704,21- diol-3,11,20-trione.

Alternatively, the 16-pregnene-3m-ol-11,20-dione 3-acetate is reacted with diazomethane to form 3u-acetoxy- 16oz,l7a-methy1eneazo-pregnane-11,20-dione which, upon heating to an elevated temperature of about 180 C., is converted to 3a-acetoxy-l6-methyl-l6-pregnene-11,20-dione. The latter compound is then reacted with hydrogen peroxide under alkaline conditions to form 16oc,17ocepoxy 30c hydroxy 16/3 methyl pregnane 11,20- dione which, upon reaction with perchloric acid inaqueous dioxane solution, is converted to a mixture of 30:,17a-dihydroxy 16 methyl 15 pregnene 11,20 dione and 30,17oc dihydroxy 16 methylene pregnane 11,20 dione. This mixture is reacted, without separation, with hydrogen in the presence of palladium catalyst to give a mixture of 3a,17a-dihydroxy-16a-methyl-pregnane-ll, ZO-dione and 3a,17a-dihydroxy-16B-methyl-pregnane-11, 20-dione, which is chromatographed on activated mag nesium silicate using chloroform, and mixtures of chloroform and methanol as diluting solvents to give both the 16w and 16B-methyl isomers in substantially pure form. The 3a,17adihydroxy-16fi-methyl-pregn-ane-11,20-dione thus obtained is then reacted with bromine in chloroform solution to give 21-bromo-3a,17a-dihydroxy-16B-methyl1 pregnane-11,20-dione which is, in turn, reacted with anhydrous potassium acetate in acetone containing potassium iodide and acetic acid catalyst to give 3a,17oc,Zl-trihydroxy 16,6 methyl pregnane 11,20 dione 21 acetate. The last-named compound is reacted with N-bromosuccinimide to form 17a,21-dihydroxy-16fi-methyl-pregnane-3,11,20-trione 2l-acetate which is reacted with bromine in chloroform-acetic acid solution to form the 4-bromo derivative which is reacted With a dehydrohalogenating agent to form 16fi-methyl-4pregnene-l7u, 21-diol-3,l1,20-trione 21-acetate, which is hydrolyzed by reaction with dilute aqueous methanolic potassium bicarbonate solution to form 16fi-methyl-4-pregnene-17a, 21-diol-3 ,11,20-trione.

The reaction between the 16-methyl-4-pregnene-17a, 21-dio1-3,11,20 trione compound, i.e. the l6a-methyl-4- pregnene-17a,21-diol-3,11,20-trione or I6 S-methyI-4-pregnene-17a,21-diol-3,11,20-trione, and formaldehyde to form the corresponding bismethylene-dioxy derivative is conveniently conducted by intimately mixing together a solution of the steroid in an organic solvent, preferably a halogenated hydrocarbon solvent such as chloroform, and a solution of formaldehyde in an aqueous mineral acid such as aqueous hydrochloric acid. The reaction is ordi narily carried out at substantially room temperature under which conditions the formation of the bismethylenedioxy derivative is substantially complete in about 2-73 days. The non-aqueous layer is recovered, washed with an aqueous alkaline solution until neutral, dried and evaporated to give the corresponding 17u,20,20,21-bis methylenedioxy-l6-methyl-4-pregnene-3,1l-dione, i.e. 20,20,21 bismethylenedioxy 16a methyl 4 pregnene 3,11 dione or 17a,20,20,21 bismethylenedioxy 16,9 methyl 4 pregnene 3,11 dione.

The reaction between ethylene glycol and this bismeth: ylenedioxy derivative (irrespective of the configuration of the 16-methyl substituent) is carried out by heating the reactants together in solution in a hydrocarbon solvent such as benzene in the presence of an acidic catalyst such as p-toluenesulfon-ic acid. Using benzene as the solvent,- the reaction is conveniently conducted by heating the re'- action solution under reflux for a period of about 15 hours, following which the cooled reaction solution is washed with an aqueous alkaline solution, dried and evaporated to give the 3-ethylenedioxy-l7u,20,20,2l-bismethylenedioxy-l6-methyl-5-pregnene-ll-one, more particularly 3-ethylenedioxy- 1'7u,20,20,21 bismethylenedioxy- 16a-methyl pregnene-ll-one; 3-ethylenedioxyi7a,20, 20,21 bismethylenedioxy-lfiji-methyl-S-pregnene-ll-one. This 3-ethylenedi0xy derivative can be purified if desired by chromatography on acid-washed alumina. While the reaction details, which are set forth hereinbelow for the conversion of this 3-ethy1enedioxy-l7a,20,20,2l-bisrnethy1enedi0xy-lG-methyI-S-pregnene-ll-one to the 6,16-dimethyl ll oxygenated 1,4 pregnadiene-l7a,2l-diol- 3,20-dione compounds, are applied to the treatment and production of compounds in which the l6-methyl grouping is in the alpha position, they are equally applicable for the treatment of, and result in the production of, the corresponding l6fl-methyl compounds.

'Ihe 3-ethylenedioxy-l7a,20,20,21-bismethylenedioxy- 16a-methy1-5-pregnene-ll-one is reacted with perbenzoic acid or perphthalic acid in the presence of a liquid medium, for example a hydrocarbon solvent such as benzene. The reaction is conveniently conducted at room temperature under which conditions the reaction is ordinarily substantially complete in about 2 days. The reaction solution is decanted from a portion of the epoxide product, and the remaining epoxide is recovered from the neutralized and dried reaction solution by evaporation. The epoxide product, that is the 3-ethylenedioxy-l7a,20, 20,2 l-bisrnethylenedioxy 5,6 epoxy 16a methyl-pregnane-ll-one, is dissolved in substantially anhydrous formic acid, and the resulting solution allowed to stand at room temperature for a period of about 2-6 hours. The reaction solution is then poured into water and the aqueous mixture is extracted with a halogenated hydrocarbon solvent such as chloroform; The chloroform extract is neutralized, dried and evaporated to give a mixture of l7u,20,20,2l bismethylenedioxy 160a methyl-6-formyloxy-pregnane-5-ol-3,l l-dione and l7a,20,20,2l-bismethylenedioxy 16oz methyl-5-formyloxy-pregnane+6-ol-3,11- dione.

The reaction between the mixed formyloxy derivatives and the alkali hydroxide solution is conveniently conducted by dissolving the formyloxy compounds in a lower alkanol, adding to this solution an aqueous solution of alkali metal hydroxide such as potassium hydroxide, and heating the resulting aqueous alcoholic alkaline solution under reflux and in the presence of a nitrogen atmosphere for a period of approximately onehalf hour. The reaction solution is cooled, neutralized with an organic acid such as acetic acid, and evaporated to small volume. The aqueous concentrate is diluted with water, and the aqueous mixture is extracted with a halogenated hydrocarbon solvent such as chloroform. The non-aqueous extract is neutralized, dried and evaporated to give 17a,20,20,21 bismethylenedioxy 16a methyl-allopregnane 3,6,11- trione, which can be purified, if desired, by crystallization from a lower alkanol such as methanol.

The reaction between the 17a,20,20,2l-bismethylenedioxy-l6a-methyl-allopregnane-3,6,1l-trione and butanone dioxolane is conveniently conducted by heating the reactants together under reflux in the presence of an acidic catalyst such as p-toluenesulfonic acid under which conditions the reaction is ordinarily complete in about 10 minutes. The reaction mixture is cooled, diluted with chloroform and the resulting chloroform solution is washed with bicarbonate, dried and evaporated to give 3-ethylenedioxy 17a,20,20,2l bismethylenedioxy 160crnethyl-allopregnane-6,1l-dione, which can be purified, if desired, by crystallization from ethyl acetate.

A solution of this 3-ethylenedioxy-17a,20,20,2l-bis methylenedioxy 16oz methyl-allopregnane-6,ll-dione in benzene is then added with stirring, over a ten-minute period, to asolution of methyl magnesiurniodide in diethyl ether. The resulting mixture is allowed to stir for about one-half hour additional time, Water is added followed by additional benzene, and'the organic layer is recovered, dried and evaporated to give the corresponding S-ethylenedioxy l7u,20,20,2l-bismethylenedioxy-fia, 1fia-dimethyl-allopregnane-6,6-01-l l-one.

The reaction between the 3-ethylenedioxy-17a,20,20,21- bismethylenedioxy 6a,l6oc dimethyl-allopregnane-QB-olll-one and thionyl chloride, when the latter is used as the dehydrating agent, is conveniently conducted by adding a solution of thionyl chloride in anhydrous pyridine drop-wise to an anhydrous pyridine solution of the steroid, while maintaining the temperature of the solution at approximately 40 C. The resulting solution is stirred for an additional one-half hour period, cooled, poured into ice water, and the aqueous mixture extracted with a halogenated hydrocarbon solvent such as chloroform. The chloroform layer is neutralized, dried, and evaporated to a crude product which is purified by chromatography over alumina to give S-ethylenedioxy-17e,20,20,21- bismethylenedioxy-6,16a-dimethyl-5-pregnene-1 l-one.

The 3-ethylenedioxy-17a,20,20,2l-bismethylenedioxy-ti, l6a-dimethyl-5-pregnene-ll-one is dissolved in anhydrous acetone containing p-toluenesulfonic acid monohydrate, and the resulting solution is allowed to stand at room temperature for a period of about 15 hours. The resulting solution is then poured into water, the aqueous mixture is extracted with a halogenated hydrocarbon solvent such as chloroform. The chloroform solution is neutralized, dried and evaporated to give 17a,20,20,2l-bismethylenedioxy-6a,l6a-dimethyl 4-pregnene-3,ll-dione. Similarly, when 3-ethylenedioxy l7d,20,20,2l-bismethylenedioxy- 16,8-methyl-5-pregnene-l l-one is used in the reaction with perbenzoic or perphthalic acid described hereinabove, instead of the 3-ethylenedioxy-17a.,20,20,2l-bisrnethylenedioxy-l6a-methyl-5-pregnene-ll-one therein employed, and the products treated in accordance with the foregoing procedures, there is obtained l7e,20,20,2l-bismethylenedioxy-6a,l65-dimethyl-4-pregnene-3,1l-dione.

In the alternate procedure for the production of the corresponding 6,6,16-dirnethyl compounds, the reaction between the 3-ethylenedioxy-17u,20,20,2l-bismethylenedioxy-5,6-epoxy-l6-methyl-pregnane-1l-one, more particularly the 3-ethylenedioxy-17a,20,20,2l-bismethylenedioxy-S,'6-epoxy-16a-methyl-pregnane-l l-one; 3-ethylenedioxy 17m,20,20,21 bismethylenedioxy 5,6 epoxy-16,8- methyl-pregnane-ll-one and the methyl magnesium bromide is conveniently conducted by heating the reactants together in benzene solution under nitrogen at a temperature of about C. for a period of about 5 hours. The reaction mixture is cooled, aqueous ammonium chloride solution is slowly added, and the mixture shaken; the henzene layer is separated, dried and evaporated to dryness to give the corresponding 17a,20,20,2l-bismethylenedioxy- 6;8,16-dirnethyl 3 ethylenedioxy-allopregnane-Swol-11- one.

The hydrolysis of the B-ethyIenedioXy-grouping is ordinarily carried out by heating the 17e,20,20,21-bismethylenedioxy-6;8,16-dimethyl 3-ethylenedioxy-allopregnane- Sm-ol-ll-one with a dilute aqueous alcoholic solution of a mineral acid, preferably a dilute aqueous methanolic solution of sulfuric acid, for a period of about one-half to one hour under reflux in a nitrogen atmosphere. The reaction mixture is cooled, aqueous sodium carbonate solution is added slowly, the mixture is shaken, and the material which precipitates is recovered by filtration and dried to give the corresponding l7a,20,20,2l-bismethylenedioxy- 6,8, l 6-dimethyl-allopregnane-3 ,l l-dione-S a-Ol.

The dehydration of the resulting allopregnane-Sa-ol to the corresponding 4-pregnene is preferably conducted by heating the 17e,20,20,2l-bismethylenedioxy-6B,l6-dimethyl-allopregnane-3,ll-dione-5ot-ol with an 0.25% solution of potassium hydroxide in methanol at reflux temperature tion mixture is diluted with water, the aqueous mixture is extracted with ethyl acetate, and the ethyl acetate extract is evaporated to dryness to give a mixture which is separated by chromatography on activated alumina followed by elution with mixtures of benzene-ether to give, as the more-ether soluble component, l7u,20,20,2l-bismethylenedioxy-6fi,16-dimethyl-4-pregnene-3,ll-dione, more particularly 17a,20,2(),2l-bismethylenedioxy-6B,l6a-dimethyl-4- pregnene-3,11-dione; 17a,2(),20,2l-bismethylenedioxy-Gfi, 16/3-dimethyl-4-pregnene-3.,l l-dione.

The reaction between the l7oz,20,20,2l-bismethylenedioxy-6,l6-dimethyl-4-pregnene-3,1l-dione, more particularly the 17a,20,20,21-bismethylenedioxy-6a,16a-dimethyl-4- pregnene-3,ll-dione; 17a,20,20,21-bismethy1enedioxy-6a, l6B-dimethyl-4-pregnene 3,11 dione; 170:,2O,20,21 bismethylenedioxy-6fi,l6ot dimethyl-4-pregnene-3,1l-dione; l7oc,2(),20,21 bismethylenedioxy-6B,16u-dimethyl-4-pregnene-3,ll-dione and the aqueous organic acid hydrolyzing agent, more particularly an aqueous acetic acid solution, is carried out by heating the reactants together at about 70 C. in a nitrogen atmosphere for a period of about 8 hours. The hydrolysis solution is evaporated to give 6,16-dimethyl-4-pregnene-l7u,21-diol-3,11,20-trione, which is then reacted with an acylating agent, e.g. acetic anhydride and pyridine to form the corresponding 6,16-dimethyl-4-preg- I1Il6-170L,21-dlOl-3,11,20-tfl0116 21-acy1ate.

The reaction between the hereinabove mentioned 3-ethylenedioxy-17a,20,20,21-bismethylenedioxy-6,16-dimethyl- -pregnene-11-one compound and lithium aluminium hydride is carried out by adding a solution of the steroid in a hydrocarbon solvent such as benzene to an ethereal suspension of the lithium aluminum hydride and heating the resulting suspension under reflux for a period of about 4 hours. Ethyl acetate is added to the cooled reaction mixture to destroy excess lithium aluminum hydride, water is then added, and the ethereal layer is recovered, dried and evaporated to give a crude crystalline material which is purified by chromatography over acid-washed alumina to give 3-ethylenedioxy-l7u,20,20,21-bismethylenedioxy- 6,l6-dimethyl-5-pregnene-11,8-ol, more particularly 3-ethylenedioxy-170,2O,20,2l-bismethylenedioxy-6,l6a-dimethyl 5 pregnene-llfi-ol; 3-ethylenedioxy-17u,20,20,21-bismethylenedioxy 6,l6B-dimethyl-5-pregnene-11,5-01. The reaction of the 3-ethylenedioxy-17a,20,20,21-bismethylenedioxy-6,l6-dimethyl-5-pregnene-113-01 and p-toluenesulfonic monohydrate in acetone is conducted in the same manner as described hereinabove in connection with the corresponding 3-ethylenedioxy-17a,20,20,21-bismethylenedioxy-6,16u-dimethyl-5-pregnene-1l-one to produce 17oz, 20,20,21-bismethylenedioxy 6a,l6-dimethyl-4-pregnene- 1l5-ol-3-one, more particularly 17a,20,20,21-bismethylenedioxy-6a,l6u-dimethyl-4 pregnene-llB-ol-S-one; 17oz, 20,20,21-bismethylenedioxy-6a,16p dimethyl-4-pregnene- 11 3-ol-3-one.

Similarly, when 17,20,20,21-bismethylenedioxy-6B,16- dimethyl-3-ethylenedioxy-allopregnane-5ot-ol-1l-one, more particularly 17a,20,20,21 bismethylenedioxy 6,3,160c-Climethyl-3-ethylenedioxy-allopregnane-Sa-ol-1l-one; 17a, 20,20,2 1 bismethylenedioxy 66,165 dimethyl 3- ethylenedioxy-allopregnane-5a-ol-ll-one is reacted with lithium aluminum hydride in accordance with the foregoing procedure and the resulting 115-01 compound hydrolyzed by reaction with dilute aqueous methanolic sulfuric acid as previously described for the hydrolysis of 17oc,20,20,21 bismethylenedioxy 65,16 dimethyl 3 ethylenedioxy-allopregnane-5a-ol-1l-one, there is obtained the corresponding 17a,20,20,2l-bismethylenedioxy-6fi,16- dimethyl-allopregnane-S 11,1 1 fi-diol-3-one. Dehydration of the latter compound with 0.25% methanolic potassium hydroxide solution, in accordance with the method previously described for the dehydration of the allopregnane- 5a-ol-ll-one compound, followed by chromatographic separation of the reaction mixture, results in the production of the corresponding 17a,20,20,2l-bismethylenedioxy-6/3,l6-dimethyl-4-pregnene-11,8-ol-3-one, more particularly 17oz,20,20,21 bismethylenedioxy 65,160: di-

1Q methyl-4-pregnene-1lfi-ol-3-one; 17a,20,20,21-bismethy1- enedioxy-6B,1613-dimethyl-4-pregnene-1 lfi-ol-one.

The 17a,20,20,21 bismethylenedioxy-6,16-dimethyl-4- pregnene-l lfl-ol-Z-one, more particularly 17a,20,20,21-bi8- methylenedioxy-6u,16a-dimethyl-4-pregnene-115-01-3-one; 17a,20,20,21 bismethylenedioxy 6a,16fi dimethyl-4- pregnene-11fl-o1-3-one; 17ot,2(),20,2l-bismethylenedioxy- 65,l6a-dimethyl-4-pregnene-l1fi-ol-3-one; 17u,20,20,21- bismethylenedioxy 6,8,16/5' dimethyl 4 pregnene- -ol-3-one is then reacted with an aqueous organic acid hydrolyzing agent such as aqueous acetic acid in the same manner as that set forth hereinabove in connection with the hydrolysis of the 17a,20,20,2l-bismethylenedie oxy-6oc,l6adimethyl-4-pregnene 3,11 dione compound, thereby forming the corresponding 6,16-dimethyl-4-pregnene-l1 5,l7a,21-triol-3,20-dione, which is reacted with an acylating agent, e.g. acetic anhydride, propionic anhydride, and the like, in pyridine to form the corresponding 2l-acylate such as 6e,16a-dimethyl-4-pregnene-11,8,17oz, 21-triol-3,20-dione ZI-acetate; 6a,16fi-dimethyl-4-pregnene-1 1B, l70c,21l1l0l-3 ,ZO-dione 21-propionate; 6B,16adlmethyl-4-pregnene-l 1B,17a,21-triol-3,20-dione 2l-acetate; 6,8,16/3 dimethyl 4 pregnene l1,8,17x,21 triol 3,20- dione 21-butyrate, and the like.

While the reaction details, which are set forth hereinbelow for the introduction of the 9a-fluoro substituent via the A 9oc-bIOH10; and 9,11-epoxide intermediates, and for the introduction of the A double bond microbiologically using Schizomycetes, or chemically using selenium dioxide, are applied to the treatment and production of 6ct,16ot-dimethyl compounds, these reaction details are equally applicable for the treatment of, and result in the production of, the other 6,16-dimethyl compounds, that is the 6a,16B-dimethyl compounds; the 6fi,l6ot-dimethyl compounds; and the 6,6,16,8-dimethyl compounds.

The reaction between the 604,16ct-dimethyl-4-pregnene- 11,8,17a,2*1-triol-3,20-dione 21-acylate and phosphorous oxychloride or methane sulfonyl chloride is conducted by bringing the reactants together in pyridine solution at room temperature and allowing the solution to stand at that temperature for a period of about 15 hours. The pyridine reaction solution is evaporated in vacuo to small volume, diluted with water and extracted with a water-immiscible organic solvent such as ethyl acetate. Evaporation of the washed and dried ethyl acetate solution gives the corresponding 60, l6u-dimethyl-4,9 l 1 -pregnadiene- 17u,21-diol-3,20-dione 21-acylate which can be purified if desired by crystallization from an organic solvent such as ethyl acetate-ether.

The addition of hypobromous acid to the A -double bond of this 6a,16a-dimethyl-4,9(11)-pregnadiene-17a,21- diol-3,20-dione 21-acylate is conveniently carried out by adding an aqueous solution of perchloric acid to a suspension of the steroid compound and N-bromo-succinimide in aqueous dioxane while maintaining the temperature of the mixture below about 15 C. The reaction mixture is allowed to stand at 15 C. for about 2-3 hours, the yellow solution is treated with allyl alcohol to discharge excess N-bromosuccinimide, and the resulting solution is evaporated in vacuo to small volume. The concentrated solution is diluted with Water and the aqueous mixture extracted with a water-immiscible organic solvent such as ethyl acetate. The washed and dried ethyl acetate extract is evaporated to dryness to give the corresponding 9a-bromo-6a,16a-dimethyl-4-preg nene-l1B,17a,21-triol-3,20-dione 21-acylate which can be purified, if desired, by crystallization from an organic solvent such as ethyl acetate-ether.

The reaction of this 9,11-brornohydrin with potassium acetate is carried out by heating the reactants together in ethanol under reflux for a period of about two hours. The cooled reaction mixture is evaporated to small volume in vacuo, the concentrated solution is diluted with water, and the aqueous mixture is extracted with ethyl acetate. The washed and dried ethyl acetate extract is evaporated '11 in dryness in vacuo to give the corresponding 6a,16m dimethyl 9,11 epoxy 4 pregnene 1701,21 diol- 3,20-dione 2l-acylate which is purified by crystallization from ethyl acetate-ether.

The reaction between this 9,11-epoxide and hydrogen fluoride is conveniently conducted by bringing the reactants together in cold alcohol-free chloroform and allowing the resulting mixture to stand at C. for about two hours. A cold aqueous solution of sodium acetate is added, and the resulting mixture is agitated vigorously. The layers are separated, and the chloroform layer is Washed, dried and evaporated to dryness in vacuo to give 6a,16a-dimethyl-9a-fiuoro-4-pregnene-l 118,l7a,21-tri- .ol-3,20-dione 2l-acylate which can be purified by crystallization from acetone-petroleum ether.

'This 16a,l6u dimethyl-9u-fluoro-4-pregnene-l 1 3,170, 2 1-triol-3,20-dione 21-acylate is then dissolved in a mixture of benzene and dilute methanolic potassium hydroxide and the mixture allowed to stand at room temperature for about minutes. The reaction mixture is acidified with acetic acid, the solvents are evaporated in vacuo :and the residual material is crystallized from ethyl acetats-ether to' give 6a,l6ot-dimethyl-9a-fiuoro-4-pregnene- 11 8, l7oc,2 1-triol-3,20-dione.

The oxidation of the llfi-hydroxyl group in the 60s, l6oz-dimethyl9a-fiuoro 4 pregnene-l1,3,l7o,21-tri0l-3, .ZO-dione ZI-acylate is conducted by dissolving the steroid ester in pyridine and adding the solution to the complex formed by adding 1 part of chromium trioxide to 5 parts 30f pyridine. The reactants are mixed thoroughly and the mixture allowed to stand at room temperature about hours. The reaction mixture is poured into water and the aqueous mixture is extracted with ether and then with ethyl acetate. The combined organic extracts are Washed, dried and evaporated to dryness in vacuo to give 6a,l6a-dimethyl-9a-fiuoro-4-pregnene-l7a,2l-diol-3, 11,20-trione Zl-acylate which can be purified by crystallization from ethyl. acetate-ethen This 21-ester is hydrolyzed with methanolic potassium hydroxide-benzene, in the same manner as described hereinabove in connection with the hydrolysis of 60c,160c-dim6ll1yl-9oc-fill0rO-4- pregnene-l1/8,l7u,2l-triol-3,20dione Zl-acylate, to produce 6a,l6a-dimethyl-9a-fluoro-4-pregnene-17oc,21-diol-3, -1 1,20-trione.

The microbiological A dehydrogenation of these 6oz,

-IGot-dimethyl-Il-oxygenated 4 pregnene-17u,21-diol-3, -dione compounds, e.g. 605,16u-dimethyl-4-pregnene 17oz,2ldiol-3,l1,20-trione; i6a,l6ot dimethyl-4-pregnene- 11fi,l7a,2l-triol-3,20-dione; 6a,l6m-dimethyl-9a-fluoro-4- pregnene-l l5,l7ot,21-triol-3,20-dione; Zl-esters of these -6a,160c dimethyl-l l-oxygenated-4-pregnene-l71,2l-diol- :.3,20-dione compounds, and the like, is conveniently conducted utilizing Schizomycetes microorganisms. This dehydrogenationreaction is eifected by contacting the steroid compound with the Schizomycetes microorganisms themselves or, if preferred, with enzyme systems of Schizomycetes microorganisms whereby the hydrogen attached to the C-1 and C-2 carbon atoms is selectively removed to produce the corresponding A -steroid substantially uncontaminated by unwanted products. When the 6oz,16u-dimethyl-1 1-oxygenated-4-pregnene-17,2l-di- .ol-3,20-dione compound is thus subjected to the dehydrogenating activity of Schizomycetes microorganisms, the corresponding 60:,16a dimethyl-l1-oxygenated-1,4-preg- .nadiene-l7u,2l-diol-3,20-dione compound is obtained directly and in high yield. This microbiological A dehydrogenation procedure is ordinarily carried out by adding the 60:, l Ga-dimethyl- 1 1-oxygenated-4-pregnene-1 7uc,2 l-diol-3,20-dione compound as a solid, or as a solution in a solvent as for example a dialkyl ketone such as acetone, a dialkyl-formamide such as dirnethyl-formamide, and :the like, under sterile conditions to a culture of the microorganism in a nutrient medium and agitating the resultingmixture thereby bringing about growth of the microorganism and dehydrogenation of the steroid com-- pound. The steroid can be added at the time the nutrient medium is inoculated with the culture of Schizomycetes microorganisms or, alternatively, may be added to an established culture. Instead of adding the steroid 5 compound to the established culture in the nutrient medium, the cell growth from such established culture may be filtered from the broth, washed with distilled water, then suspended in buffered aqueous solution containing the 6u,16m-dimethyl-11-oxygenated-4-pregnene-l7a,2l-diol-3,20-dione compound, and the resulting mixture agitated thereby effecting dehydrogenation of the steroid .compound to form the corresponding 6a,l6udimethyl-1l- .oxygenated-l,4-pregnadiene-l7a,21-diol-3,20-dione. The latter is more readily recovered from this medium than from the mixture obtained when the steroid is incubated with the microorganisms in the original nutrient medium. Alternatively, the 6u,16a-dimethyl-l1-oxygenated-4-pregnene-l7u,21-diol-3,20-dione compound may be contacted with dehydrogenating enzyme preparations from the growth of Schizomycetes microorganisms.

The nutrient mediums used in carrying out this bacteriological dehydrogenation are those ordinarily utilized in the propagation of Schizomycetes microorganisms. The usual nutrients include a nitrogen and carbon source, inorganic salts and growth factors when required. The carbon can be provided by compounds such as acetates, lactates, and the like. The nitrogen can be provided by an ammonium salt, amino acids, or proteins such as soy .beans, oats, yeast, yeast extracts, tryptic digest of casein, meat extract, blood meal, protein meat and bone scrap, salmon meal, fish meals, fish solubles, distillers solubles, and the'like. If desired, the Schizomycetes microorganisms can be propagated using proteins (or amino acids) without any carbohydrate being present in the medium, in which case the proteins 'or amino acids serve as the source of both the carbon and nitrogen required by the microorganisms.

While, as noted hereinabove, the dehydrogenation of the 60:,160: dimethyl-1l-oxygenated-4-pregnene-l70:,21- .diol-3,20-dione compound may be carried out using dehydrogenating enzyme preparations from the growth of .Schizomycetes microorganisms, or by contacting the steroid compound with a suspension of an established culture in distilled water, it is ordinarily preferred to add the 6a,16m dimethyl-ll-oxygenated-4-pregnene-1711,21- diol-3,20-dione compound to a nutrient medium containing a 24-hour growth of Schizomycetes microorganisms. T he proportion of steroid compound which may be added .to the medium varies depending upon the particular substrate being dehydrogenated, but it is ordinarily preferred to employ a concentration of about 0.005% to 0.2% of .6a,l6a dimethyl-l I-oXygenatedl-pregnene-17,2l-diol- 3,20-dione compound, although higher or lower concentrations may be employed, if desired. The culture con- 55-taining the added steroid compound is then incubated,

preferably with agitation and aeration for an additional period which ordinarily varies between about 10 hours and hours, although shorter or longer fermentation times may be advantageous for the dehydrogenation of 0 particular substrates. In view of the fact that prolonged fermentations may result in destruction of a portion of the A -dehydrogenated steroid product, it is ordinarily preferred to employ a fermentation time of about 10 hours to 24 hours which, depending upon the steroid substrate, has been found to result in maximal yields of the A -dehydrogenated steroid product.

" Aftercompletion of the dehydrogenation process, the

160: dimethyl-ll-oxygenated-l,4-pregnadiene-l7a,2l-

diol-3,20-dione product is conveniently recovered from the fermented broth by extraction with a water-immiscible solvent as for example a chlorinated hydrocarbon such "as chloroform, a ketone such as methyl isobutyl ketonc,

an alkyl alkanoate such as ethyl acetate, and the like.

The extract of A -dehydrogenated steroid product and any unreacted starting material which may be present is con- -11 oxygenated 4-pregnene-17a,21-diol-3,20-dione compound is reacted with selenium dioxide thereby effecting ring A dehydrogenation to form the corresponding 6,16- dimethyl ll oxygenated 1,4-pregnadiene-17,21-diol- 3,20-dione compound. This selenium dioxide dehydrogenation procedure is conveniently conducted by bringing the 6,16-dimethyl-ll-oxygenated-4-pregnene-17,21- diol-3,20-dione compound, and selenium dioxide together in the presence of an organic solvent such as for example dioxane, an alcohol solvent such as t-butanol, etc., and heating the mixture at an elevated temperature. When t-butanol is used as the solvent, it is ordinarily preferred to carry out this reaction at the boiling point of the solvent, under which conditions the reaction is ordinarily complete in about fifteen hours. The reaction mixture is ordinarily filtered, thereby removing metallic selenium, and the filtered solution is evaporated to dryness in vacuo to give the desired 6,16-dirnethyl-1loxygenated-l,4-pregnadiene-170;,21 diol-3,20-dione com pound. The crude material obtained in this way is conveniently purified by paper strip chromatography in accordance with the procedure outlined hereinabove in connection with the purification of the 6,16-dimethyl-lloxygenated-1,4-pregnadiene l7oc,2l-diol-3-20-dione compound produced by microbiological dehydrogenation.

The following examples illustrate methods of carrying out the present invention but it is to' be understood that these examples are given for purposes of illustration and not of limitation.

Example 1 grams of 16u-methyl4-pregnene-l7a,2l-diol-3,11, 20-trione are dissolved in 400 cc. of chloroform. To this solution is added a mixture of 100 cc. of concentrated aqueous hydrochloric acid and 100 cc. of 37% aqueous formaldehyde solution. The resulting mixture is stirred for a period of approximately 3 days at room temperature. The chloroform layer is separated, washed with sodium bicarbonate solution until neutral, then with water, dried over anhydrous sodium sulfate, and evaporated to dryness in vacuo. The residual crude material is purified by crystallization from a mixture of methylene chloride and methanol to give l7oc,20,20,2lbismethylenedioxy-16a-methyl-4-pregnene-3, 1 l-dione.

10 grams of 17a,2(),20,21 bismethylenedioxy 16amethyl-4-pregnene-3,1l-dione are dissolved in 500 cc. of benzene, to the solution is added 25 cc. ethylene glycol and 1 gram of p-toluenesulfonic acid, and the resulting mixture is heated under reflux for a period of approximately hours. The reaction solution is cooled, washed With aqueous sodium bicarbonate solution, dried over anhydrous sodium sulfate, and evaporated to dryness in vacuo. The residual crude material is adsorbed from its solution in benezene on 250 g. of acid-washed alumina, and the resulting adsorbate is eluted utilizing mixtures of ether-petroleum ether to give 3-ethylenedioxy l7a,20,20,21 bismethylenedioxy 16oz methyl-5- pregnene-l 1-one.

A mixture of 10 grams of S-ethylenedioxy-17bz,20,20, 2l-bismethylenedioxy-l6a-rnethyl-5-pregnene-ll-one, 4.3 g. perbenzoic acid and 230 cc. benezene is allowed to stand at room temperature for a period of about 2 days. A portion of the epoxide product, which precipitates, is separated from the reaction solution by decantation, and the decanted solution is washed with satured aqueous sodium bicarbonate solution and then dried over anhydrous sodium sulfate. The dried solution is then evaporated in vacuo, and the residual epoxide product is combined with the epoxide product originally separated from the reaction mixture to give 3-ethylenedioxy- 17m,20,20,21 bismethylenedioxyl6a-methyl-5,6-epoxypregnane-1 l-one.

The latter product is dissolved in 200 cc. of substantially anhydrous formic acid and the resulting solution is allowed to stand at room temperature for a period of about 2 /2 hours; The reaction solution is poured into water, and the aqueous mixture is extracted with chloroform. The chloroform solution is washed with saturated aqueous sodium bicarbonate solution, dried over anhydrous sulfate, and evaporated in vacuo to give a mixture of 17a,20,20,21-bismethylenedioxy- 16a-methyl-6- formyloxy pregnane S-ol-3,1l-dione and 17oz,20,20,2lbismethylenedioxy 16a-methyl-5-formyloxy-pregnane-6- ol-3,11-dione.

This mixture of S-formyloxy and fi-formyloxy derivatives is dissolved in about 850 cc. of methanol, to this solution is added a solution containing about 17 grams of potassium hydroxide in cc. of water, and the resulting mixture is heated under reflux in a nitrogen atmosphere for a period of approximately /2 hour. The reaction solution is cooled, neutralized with about 23 cc. of acetic acid and evaporated in vacuo to a small volume. The concentrated solution is poured into water and the aqueous mixture is extracted with chloroform. The chloroform extract is washed with aqueous sodium bicarbonate solution, dried over anhydrous sodium sulfate, and evaporated to dryness in vacuo. The residual material is crystallized from methanol to give 17a,20,20, 21-hismethylenedioxy loot-methyl allopregnane-3,6,11- trione.

A solution containing about 5 grams of l7u,20,20,21- bismethylenedioxy 16a-methylallopregnene-3,6,11trione and 0.1 g. of p-toluenesulfonic acid in cc. butanone dioxolane is heated under reflux for a period of about 10 minutes. The reaction solution is cooled to about 05 C., diluted with chloroform, and the resulting solution is washed with equeous sodium bicarbonate solution and dried over anhydrous sodium sulfate. The resulting dried solution is evaporated to dryness in vacuo, and the residual material is slurried with ether, recovered by filtration and then dried to give a 3-ethylenedi0xy- -17a,20,20,21 bismethylenedioxy 16m methyl-allopregnane-6,l1-dione.

A solution containing about 5 grams of 3-ethylenedioxy 1704,20,Z0,21 bismethylenedioxy 16a methylallopregnane-6,1l-dione in 70 cc. of benzene is added, with stirring over a period of approximately 10 minutes, to an ethereal solution containing the methyl magnesium iodide, prepared from 5 cc. of methyl iodide and 0.5 gram of magnesium turnings, dissolved in 50 cc. of ether. The reaction mixture is allowed to stir for an additional /2 hour period, and the resulting solution is decomposed with 70 cc. of water. About cc. of benzene is added to the aqueous mixture and the layers are separated. The aqueous layer is extracted with two 100 cc.-portions of, chloroform, and the organic. layers are combined, dried over anhydrous sodium sulfate, and evaporated to dryness in vacuo to give 3-ethylenedioxy-l7u,20,20,21- bismethylenedioxy 6&1601. dimethyl allopregnane- 6/3-ol-1l-one.

A solution containing 5.5 cc. of freshly distilled thionyl chloride in 26 cc. of cold anhydrous pyridine is added dropwise, with stirring, to a solution of 5.0 g. of 3 ethylenedioxy 17a,20,20,21 bismethylenedioxy- 6a,16a-dimethyl-allopregnene-ofi-ol-ll-one in 32 cc. of anhydrous pyridine, while maintaining the temperature of the reaction mixture at approximately 40 C. The reaction solution is stirred for an additional 30-minute period following the addition of the thionyl chloride reagent, and the reaction mixture is then cooled to about 05 C. and poured into cc. ice water. The aqueous mixture is extracted with chloroform and the chloroform extract is neutralized, dried over anhydrous sodium sulfate, and evaporated to dryness in vacuo. The residual material, dissolved in benzene, is chromatographed on 150 grams of acid-Washed alumina; elution of the resulting adsorbate with mixtures of petroleum ether and ether gives 3-.ethylenedioxy-l711,20,20,2l-bisn1ethylenedioxy-6,l6a-dimethyl-5-pregnene-1l-one.

A solution containing about 5 grams of .3-ethylenedioxy -17a,20,20,21 bismethylenedioxy 6,160; dimethyl-5-pregnene-1'1-one, 500 cc. of anhydrous acetone and about 0.5 g. of p-toluenesulfonic acid monohydrate is allowed to stand for a period of about 15 hours. The reaction solution is poured into water, and the aqueous mix ture is extracted with chloroform. The chloroform solution is washed with aqueous sodium bicarbonate solution, dried over anhydrous sodium sulfate, and the chloroform solution is evaporated to dryness in vacuo. The residual material is crystallized from benzene to give 17u,20,20,21 bismethylenedioxy 60,16a dimethyl 4- pregnene-3,11-dione.

About 0.1 gram of 17a,20,20,2l-bismethylenedioxy- 6a,16a-dimethyl-4-pregnene-3,ll-dione is suspended in 18 cc. of 50% aqueous acetic acid, and the suspension is carefully purged with nitrogen, following which the suspension is heated on a steam bath under nitrogen for a period of about 8 hours and then allowed to stand at room temperature for an additional period of 15 hours. The reaction solution is evaporated to dryness in vacuo to give 6oz,16a-dimethyl-4-pregnene-17a,21-diol- 3,11,20-trione. This material is dissolved in 20 cc. of chloroform, and the chloroform solution evaporated to dryness in vacuo. To the residual dry material is added 1 cc. of pyridine and 1 cc. of acetic anhydride, and the resulting mixture is heated on a steam bath for a period of about 15 minutes. The reaction solution is cooled to about 5 C. and poured into ice Water. The aqueous suspension is extracted with chloroform, and the chloroform extract is washed with aqueous sodium bicarbonate solution, dried over anhydrous sodium sulfate, and evaporated to dryness in vacuo. The residual material, dissolved in 30 cc. benzene, is adsorbed on grams of acid-washed alumina; the adsorbate is washed wtih 50 cc. ether. The adsorbate is then eluted with 250 cc. of a 5:5 mixture of ether-chloroform, and then with 50 cc. of a 3:7 mixture of ether-chloroform. The eluates are evaporated in vacuo, and the residual material is crystallized from methanol to give 604,16ocdimethyl 4 pregnene 17a,21 diol 3,11,20 trione 21-acetate.

To a solution of 100 mg. of 6a,16a-dimethyl-4-preg- I16I1-l7u,21-diOl-3,l1,20-1110116 2l-acetate, in 6 ml. t-butanol, and 0.01 ml. of glacial acetic acid is added 70 mg, of selenium dioxide. The mixture is heated under reflux in a nitrogen atmosphere for a period of about 6 hours. The reaction solution is filtered and evaporated in vacuo. The residual material is dissolved in benzene, and the benzene solution is washed with an aqueous sodium bicarbonate solution and dried over anhydrous sodium sulfate. The benzene solution is stirred with mercury overnight, then centrifuged, and the clear benzene solution is evaporated in vacuo. The residual material thus obtained is separated using partition chromatography on diatomaceous silica (supercel) to give 6a,16tx-dimethyl-1,4-pregnadiene-17u,21-diol-3,-11, 20-trione 2l-acetate.

ExampleZ To a solution of 100 mg. of 17a,'20,20,21-bismethylenedioxy 60,160t dimethyl 4 pregnene 3,11 dione, prepared as described in Example 1 hereinabove, in 6 ml. t-butanol, and 0.01 ml. of glacial acetic acid is added 70 mg. of selenium dioxide. The mixture is heated under reflux in a nitrogen atmosphere for a period of about 6 hours. The reaction solution is filtered and evaporated in vacuo. The residual material is dissolved in benzene, and the benzene solution is washed with an aqueous sodium bicarbonate solution and dried over anhydrous sodium sulfate. The benzene solution is stirred with mercury overnight, then centrifuged, and the clear benzene solution is evaporated in vacuo. The residual material thus obtained is separated using partition chromatography on diatomaceous silica (supercel) to give 2.2 17oc,20,'20,21 bismethylenedioxy 60:,160: dimethyll,4-pregnadiene,3, 1 l-dione.

About 0.1 gram of 17a,20,20,21-bismethylenedioxy- 6a,l6a-dimethyl-1,4-pregnadiene-3,1l-dione is suspended in 18 cc. of 30% aqueous acetic acid, and the suspension is carefully purged with nitrogen, following which the suspension is heated on a steam bath under nitrogen for a period of about 8 hours and then allowed to stand at room temperature for an additional period of 15 hours. The reaction solution is evaporated to dryness in vacuo to give 6a,ldwdimethyl-1,4-pregnadiene-l7ot,21-di0l-3,l1, 20-trione. This material is dissolved in 20 cc. of chloroform, and the chloroform solution evaporated to dryness in vacuo. To the residual dry material is added 1 cc. of pyridine and 1 cc. of acetic anhydride, and the resulting mixture is heated on a steam bath for a period of about 15 minutes. The reaction solution is cooled to about O5 C. and poured into ice water. The aqueous suspension is extracted with chloroform, and the chloroform extract is washed with aqueous sodium bicarbonate solution, dried over anhydrous sodium sulfate, and evaporated to dryness in vacuo. The residual material, dissolved in 30 cc. benzene, is adsorbed on 5 grams of acidwashed alumina; the adsorbate is washed with 50 cc. ether. The adsorbate is then eluted with 250 cc. of a 5 :5 mixture of ether-chloroform, and then with 50 cc. of a 3:7 mixture of ether-chloroform. The eluates are evaporated in vacuo, and the residual material is crystallized from methanol to give 6a,16a-dimethyl-1,4-pregnadiene-17a,21-diol-3,l1,20-trione ZI-acetate.

Example 3 About 5 grams of 3-ethylenedioxy-17a,20,20,21-bismethylenedioxy-6, 1 6oL-dll1'l3tl1Yl-5 pregnene-1 l-one, which can be prepared as described in Example 1 hereinabove, is dissolved in about cc. of benzene. This solution is added to a stirred suspension containing 5 grams of lithium aluminum hydride in one liter of anhydrous ether, and the resulting suspension is heated under reflux for a period of about 4 hours. The reaction mixture is allowed to cool to about room temperature, and approximately 33 cc. of ethyl acetate is added to destroy excess lithium aluminum hydride. The resulting mixture is admixed with about cc. of water, the ethereal layer is decanted, and the aqueous layer, diluted with an additional 200 cc. of Water, is extracted with chloroform. The ether and chloroform layers are combined, and the combined organic layers are dried over anhydrous sodium sulfate and evaporated to dryness in vacuo. The residual material, dissolved in about 250 cc. of benzene, is adsorbed on about grams of acid-washed alumina, and the resulting adsorbate is eluted utilizing mixtures of petroleum ether-ether. This eluate is evaporated in vacuo, and the residual material is purified by crystallization from ether to give 3-ethylenedioxy-17a, 23,2012l-bismethylenedioxy 6,160c dimethyI-S-pregnene- A solution containing about 5 grams of 3-ethylenedioxy-17tx,20,20,21-bismethylenedioxy 6,16a-dimethyl-5- pregnene-11,801, 500 cc. of anhydrous acetone and about 0.5 g. of p-toluenesulfonic acid monohydrate is allowed to stand for a period of about 15 hours. The reaction solution is poured into water, and the aqueous mixture s extracted with chloroform. The chloroform solution 1s washed with aqueous sodium bicarbonate solution, dried over anhydrous sodium sulfate, and the chloroform solution is evaporated to dryness in vacuo. The residual materlal is crystallized from benzene to give 17a,20,20, 21-bismethy1enedioxy-6a,16a-dimethyl 4 pregnene-1118'- ol-3-one.

About 0.1 gram of 17a,20,20,2l-bismethylenedioxy- 6a,1'6u-dimethyl-4-pregnene-11fl-ol-3-one is suspended in 18 cc. of 50% aqueous acetic acid, and the suspension is carefully purged with nitrogen, following which the suspension is heated on a steam bath under nitrogen for a period of about 8 hours and then allowed to stand at ether.

room temperature'for an additional period of 15 hours. The reaction solution is evaporated to dryness in vacuo to give 6a,16a-dimethyl-4-pregnene-ll,B,l7a,2l-triol-3,20- dione. This material is dissolved in 20 cc. of chloroform, and the chloroform solution evaporated to dryness in vacuo. To the residual dry material is added 1 cc. of pyridine and 1 cc. of acetic anhydride, and the resulting mixture is heated on a steam bath for a period of about 15 minutes. The reaction solution is cooled to about 5 C. and poured into ice water. The aqueous suspension is extracted with chloroform, and the chloroform extract is Washed with aqueous sodium bicarbonate solution, dried over anhydrous sodium sulfate, and evaporated to dryness in vacuo. The residual material, dissolved in 30 cc. benzene, is adsorbed on grams of acidwashed alumina; the adsorbate is Washed with 50 cc. The adsorbate is then eluted with 250 cc. of a 5:5 mixture of ether-chloroform, and then with 50 cc. of a 3:7 mixture of ether-chloroform. The elnates are evaporated in vacuo, and the residual material is crystallized from methanol to give 6oz,l6oc-dimethyl-4-pregnenel1B,17u,2l-triol-3,20-dione 2l-acetate.

To a solution of 100 mg. of 6a,l6ot-dimethyl-4-pregnone-1TB,17a,21-triol-3 ,20-dione ZI-acetate, in 6 ml. t-butanol, and 0.01 ml. of glacial acetic acid, is added 70 mg. of selenium dioxide. The mixture is heated under reflux in a nitrogen atmosphere for a period of about 6 hours. The reaction solution is filtered and evaporated in vacuo. The residual material is dissolved in benzene, and the benzene solution is washed with an aqueous sodium bicarbonate solution and dried over anhydrous sodium sulfate. The benzene solution is stirred with mercury overnight then centrifuged and the clear benzene solution is evaporated in vacuo. The residual material thus obtained is separated using partition chromatography on diatomaceous silica (Supercel) to give 6 x,l6cc-dimethyl-1,4-pregnadiene-1113,l7a,2l-triol 3,20 dione 21- acetate.

Example 4 To a solution of 100 mg. of l7ot,20,20,2l-bismethylenedioxy-6a16a-dimethyl-4-pregnene-11,6-ol-3-one, prepared as described in Example 1 hereinabove, in 6 ml. t-butanol, and 0.01 ml. of glacial acetic acid is added 70 mg. of selenium dioxide. The mixture is heated under reilux in a nitrogen atmosphere for a period of about 6 hours. The reaction solution is filtered and evaporated in vacuo. The residual material is dissolved in benzene, and the benzene solution is washed with an aqueous sodium bicarbonate solution and dried over anhydrous sodium sulfate. The benzene solution is stirred with mercury overnight then centrifuged and the clear benzene solution in evaporated in vacuo. The residual material thus obtained is separated using partition chromatography on diatomaceous silica (Supercel) to give 17a,20,20,21-bismethylenedioxy-6a,1 ou-dimethyl-1,4-pregnadiene-1 1,8-ol-3-one.

About 0.1 gram of l7u,20,20,21-bismethylenedioxy6u, 1 fia-dimethyl-l ,4-pregnadiene-l l 18-01-3 -one is suspended in 18 cc. of 50% aqueous acetic acid, and the suspension is carefully purged with nitrogen, following which the suspension is heated on a stream bath under nitrogen for a period of about 8 hours and then allowed to stand at room temperature for an additional period of 15 hours. The reaction solution is evaporated to dryness in vacuo to give 611,160: dimethyl 1,4 pregnadiene-l1fl,l7a,2l-triol-3,20 dione. This material is dissolved in 20 cc. of chloroform, and the chloroform solution evaporated to dryness in vacuo. To the residual dry material is added 1 cc. of pyridine and 1 cc. of acetic anhydride, and the resulting mixture is heated on a steam bath for a period of about 15 minutes. The reaction solution is cooled to about 05 C. and poured into ice water. The aqueous suspension is extracted with chloroform, and the chloroform extract is washed with aqueous sodium bicarbonate solution, dried over anhydrous sodium sulfate, and evaporated to dryness in vacuo. 'The residual material, dissolved in 30 cc. benzene, is adsorbed on 5 grams of acid-washed alumina; the adsorbate is washed with 50 cc. ether. The adsorbate is then eluted with 250 cc. of a 5:5 mixture of ether-chloroform, and then with 50 cc. of a 3:7 mixture of ether-chloroform. The eluates are evaporated in vacuo, and the residual material is crystallized from methanol to give 6a,1Ga-dimethyl-1,4-pregnadiene-ll5 17q,21-triol-3,20-dione ZI-acetate.

Example 5 To a cooled solution of 436 mg. of 6a,16u-dimethyl-4- pregnene-l1,8,17a,21-triol-3,20-dione 21-acetate (which canbe prepared as described in Example 3 hereinabove) in 2.5 cc. dimethyl formamide and 2.0 ml. of dry pyridine is added 1.0 ml. of methane sulfonyl chloride, while maintaining the temperature below 0 C. The resulting mixture is allowed to Warm to room temperature, at which point a precipitate appears; the resulting mixture is then heated to a temperature of about 70100 C. for a period of about 10 minutes. About 15 ml. of water is added slowly to the reaction mixture, with stirring, and the aqueous mixture is extracted with ethyl acetate. The combined ethyl acetate extracts are washed with water, then with dilute aqueous hydrochloric acid solution, again with water, and then with a dilute aqueous sodium bicarbonate solution. The washed ethyl acetate solution is dried, and the solvent is evaporated in vacuo; the residual material is triturated with ether, and the crystalline material is recrystallized to give 60:,l6a-dimethyl-4,9411 1)-pregnadiene-17u,21-diol-3,20-dione 21- acetate. A suspension of 330 mg. of 6a,l6oz-CllmihYl-4,9 (ll)-pregnadiene-l7a, 21-diol-3,20-dione 2l-acetate and 1.8 g. of N-bromo-succinimide in a mixture of 50 ml. of dioxane and 10 ml. of Water is cooled to 10 C. Then with stirring, 10 ml. of cold 1.0 N aqueous perchloric acid is added. The temperature of the reaction mixture is allowed to rise to 15 C. and is maintained at this point for about two-and-one-half hours during which time the solid material slowly dissolves. The resulting yellow solu tion is treated with 1.0 ml. of allyl alcohol to discharge the color and the remaining N-bromo-succinimide, and the remaining solution is evaporated to a small volume in vacuo. The concentrated solution is diluted with water, and the aqueous mixture is extracted with ethyl acetate.

.The ethyl acetate extracts are washed, dried and evaporated to dryness, and the residual material is crystallized from ethyl acetate-ether to give 9wbromo-6a,l6a-dimethyl-4-pregnene-1 113,17)1-triol-3,20-dione 2l-acetate.

A solution of 210 mg. of 9a-bI'Ol'I'10-6d,16a-dll'l'lethy1-4r pregnene-llfl,17a,2l-triol-3,20-dione Zl-acetate and 240 mg. of potassium acetate in 10 ml. of absoluteethanol is heated under reflux for two hours. The reaction mixture is cooled to room temperature, evaporated in vacuo to a small volume, and diluted with water. The concentrated aqueous mixture is extracted with three portions of ethyl acetate, and the combined ethyl acetate extracts are washed with water, dried, and evaporated to dryness in vacuo. The residual material is crystallized from ethyl acetate-ether to give 6u,l6a-dimethy1-9,1l-epoxy-4-pregnenes-l7ot,2l-diol-3,20-dione 2l-acetate.

To a solution of 83 mg. of anhydrous hydrogen fluoride in 4.7 ml. of ice-cold alcohol-free chloroform is added an ice-cold solution of 416 mg. of 6c=,l6a-dimethy1-9,11 epoxy-4-pregnene-17a,21 diol-3,20-dione 2l-acetate. The solution is mixed thoroughly and maintained at 0 C. for two hours, at the end of which time 13.0 ml. of ice-cold 20% aqueous sodium acetate is added, and the resulting mixture agitated vigorously. The layers are separated, and the chloroform layer is washed with water until free of acid, dried, and the chloroform evaporated in vacuo. The residual material is crystallized from acetone-petroleum ether to give 6a,l6a-dimethyl-9a-fluoro-4-pregnene-11B,17a,2l-triol 3,20 dione 2l-acetate. Fifty milligrams of 6a.,16a-dimethyl-9oc-fluoro-4-pregnene 2:5 llfi,l7a,2l-triol-3,20-dione 21-acetate is dissolved in a mixture of 1.0 cc. of benzene and 1.0 cc. of 1 N methanolic potassium hydroxide, and the solution is allowed to stand at room temperature for a period of about 10 minutes. The solution is then acidified with acetic acid, the benzene is evaporated in vacuo, and the residual material is purified by crystallization to give 6a,l6a-dimethyl 9a-fluoro-4-pregnene-l 1B,l70z,21-t1l0l-3,20-dl0l1.

To a solution of 100 mg. of 6a,IGa-dlmcthYl-9rx-fil1010- 4-pregnene-l1,8,l7a,21-triol-3,20-dione 21-acetate, in 6 ml. t-butanol, and 0.01 ml. of glacial acetic acid is added 70 mg. of selenium dioxide. The mixture is heated under reflux in a nitrogen atmosphere for a period of about 60 hours. The reaction solution is filtered and evaporated in vacuo. The residual material is dissolved in benzene, and the benzene solution is washed with an aqueous sodium bicarbonate solution and dried over anhydrous sodium sulfate. The benzene solution is stirred with mercury overnight, then centrifuged, and the clear benzene solution is evaporated in vacuo. The residual material thus obtained is separated, using partition chromatography on diatomaceous silica (Supercel), to give 6a,l6a-dimethyl-9a-fiuoro 1,4 pregnadiene-llfi,17a,2ltriol-3,20-dione 2l-acetate. This material is hydrolyzed using 1 N methanolic potassium hydroxide in benzene as described above for the hydrolysis of 6u,16a-dimethyl- 9oc-fluOIO 4-pregnene-l1,6,l7a,2l-t1iol-3,20-dione 21-acetate to give 6a,16a-dimethyl-9a-fiuoro-1,4-pregnadienel1,8,l7a,2l-triol-3,20-dione.

Example 6 A solution of 400 mg. of 6a,16a-dimethyl-9a-fluoro-4- pregnene-l1,3,17a,2l-triol-3,20-dione 2l-acetate in 4 ml. of pyridine is added to the complex formed by the addition of 400 mg. of chromium trioxide to 4 ml. of pyridine. The mixture is swirled until thoroughly mixed and then allowed to stand at room temperature overnight. The reaction mixture is poured into water, and the aqueous mixture is extracted with ether and then twice with ethyl acetate. The combined ether and ethyl acetate extracts are washed with dilute aqueous sulfuric acid at about C., and then with water until neutral. The organic solvent layer is then dried, the solvents are evaporated therefrom in vacuo, and the residual crystalline material is purified by crystallization to give 6a,l6rx-dimethyl-9afiuoro 4-pregnene-l7 a,21-diol 3,11,20-trione 21-acetate, Fifty milligrams of 6a,16u-dimethyl-9a-fiuoro-4-pregnene- 17u,2l-diol-3,l1,20-trione 21-acetate is dissolved in a mixture of 1.0 cc. of benzene and 1.0 cc. of l N methanolic potassium hydrom'de, and the solution is allowed to stand at room temperature for a period of about minutes. The solution is then acidified with acetic acid, the benzene is evaporated in vacuo, and and the residual material is purified by crystallization to give 60,160L-dlIIl6ihYl-9afiuoro-4-pregnene-170:,21-di0l-3,11,20-trione.

To a solution of 100 mg. of 6a,l6u-di1nethyl-9a-fluoro- 4-pregnene-l7a,21-diol-3,l1,20-trione 21-acetate, in 6 ml. t-butanol, and 0.01 ml. of glacial acetic acid is added 70 mg. of selenium dioxide. The mixture is heated under reflux in a nitrogen atmosphere for a period of about 60 hours. The reaction solution is filtered and evaporated in vacuo. The residual material is dissolved in benzene, and the benzene solution is washed with an aqueous sodium bicarbonate solution and dried over anhydrous sodium sulfate. The benzene solution is stirred with mercury overnight, then centrifuged, and the clear benzene solution is evaporated in vacuo. The residual material thus obtained is separated using partition chromatography on diatomaceous silica (Supercel) to give 6a,l6udimethyl-9a-finoro 1,4 pregnadiene-17a,21-diol-3,ll, 20-trione 2l-acetate. This material is hydrolyzed using 1 N methanolic potassium hydroxide in benzene as described hereinabove for the hydrolysis of 6: ,16o-dimethyl- 9a-fiuoro-4-pregnene-l7a,21-dio1-3,l1,20-trione 2 l-acetate to give 6a,16a-dimethyl-9a-fluoro-1,4-pregnadiene-l7a,21- diol-3,1 1,20-trione.

Example 7 Fifty milliliters of a nutrient medium are prepared having the following composition:

Distilled Water to make 50 ml.

This medium is adjusted to pH 6.5 with KOH, sterilized and inoculated with about 2.5 to 5 ml. of a culture of Bacillus sphaericus (ATCC 245) microorganisms, and the inoculated culture is then incubated at a temperature of 28 C., with agitation, for a 24-hour period. To the resulting culture is added a solution containing 10 mg. of 60a, 1 6u-dimethyl-4-pregnenel7a,21di0l-3,1 1,20-trione dissolved in 0.2 ml. of dimethylformamide. The culture containing the steroid compound is incubated, with agitation, for an additional period of about 24 hours at 28 C.

The fermentation broth is extracted with four 5-0 ml.- portions of ethyl acetate, and the ethyl acetate extracts are combined and evaporated in vacuo to a volume of about 5 ml. The concentrated solution is then streaked on paper chromatograms which are developed using formamide as the stationary phase and 50% benzene-5 0% chloroform as the mobile phase. After 8 hours development in a descending system, the upper bands for each chromatogram, corresponding to the A -dehydro derivative, are cut ofi, extracted with methanol, and the methanol-extracted material is again subjected to streak-paper chromatography. The upper band is again out off, thoroughly dried, extracted with methanol, and the methanol extract is evaporated to dryness in vacuo. The residual material is recrystallized to give 6a,l6u-dimethyl-l,4-pregnadiene-l 7a,2 1-diol-3 ,11-20-trione.

The 6a,l6u-dimethyl-l,4-pregnadiene-17a,21-diol-3,11, 20-trione is treated with acetic anhydride and pyridine to give the 21-acetyl derivative, which is purified by recrystallization to give substantially pure 6u,l6a-dimethyl-l,4- pregnadiene-17a,21-diol-3,l1,20-trione 21-acetate.

Example 8 Fifty milliliters of a nutrient medium are prepared having the following composition:

Cerelose g 1 Edamin g l Cornsteep liquor ml 0.25

Distilled water to make 50 ml.

This medium is adjusted to pH 6.5 with KOH, sterilized and inoculated with about 2.5 to 5 ml. of a culture of Nocardia asteroides (ATCC 9970) microorganisms, and the inoculated culture is then incubated at a temperature of 28 C., with agitation, for a 24-hour period. To the resulting culture is added a solution containing 10 mg. of 6a,l6u dimethyl-4-pregnene-llfi,17a,2l-triol-3,20-dione dissolved in 0.2 ml. of dimethylformamide. The culture containing the steroid compound is incubated, with agitation, for an additional period of about 24 hours at 28 C.

The fermentation broth is extracted with four 50 ml.- portions of ethyl acetate, and the ethyl acetate extracts are combined and evaporated in vacuo to a volume of about 5 ml. The concentrated solution is then streaked on paper chromatograms which are developed using formamide as the stationary phase and 50% benzene- 50% chloroform as the mobile phase. The upper bands for each chromatogram corresponding to the A -dehydro derivative are cut off, extracted with methanol, and the methanol-extracted material is again subjected to streakpaper chromatography. The upper band is again out off, thoroughly dried, extracted with methanol, and the methanol extract is evaporated to dryness in vacuo. The residual material is recrystallized to give 60:,l6oc-dill16thYl 1,4-pregnadiened 1,8,l7u-21-triol-3,20 dione.

The 60:, l6zx-dimethyl-1,4-pregnadiene-1 1p, 17a,21-triol- Example 10 The fermentation procedures of Examples 7, 8 and 9 are repeated but using, in place of the microorganisms and the 6a,l6ot-dimethyl-l1-oxygenated-4-pregnene-17a,

Example 9- 2l-diol-3,20-dione starting material employed in those tr 'n n steroid startin -F1fty mrlhliters of a nutrient medium are prepared havexamples l lincroollgamsm 3 a1 5 d g ing the following compositioncompounds indicated in the table hereinbelow. The resulting fermentation broths are treated in accordance with Cerelose 8 1 the isolation methods described in Examples 7, 8 and 9 Fdamm 1 10 to give, for the particular microorganism strain and ste- C?n 1steeP hquor roid substrate used, the 6a,l6u-dimethyl-1l-oxygenated- Dlstlned Water to make 50 1,4-pregnadiene-l7u,21-diol-3,20-dione indicated in the This medium is adjusted to pH 6.5 with KOH, sterilized f li Wing table:

Expt. 6a,16a-dimethyl-ll-oxygenated- No. Substrate Microorganism 1,4-preg'l1adiene-17a,21-di0l- 3,20-dione product 1 6a,lfia-dimethyl-Qa-fluoro-- Bacillus sphaericus 611,160:-dirnethyl-Qa-fluoro-1,4-

preg'nene-Ua,2l-diol-3,11,20- ATC C-7055 pregnadiene-Ua,21-d1ol-3,l1,20- trione 21-acetate. V I tr1one.

3 GaJGaAimethyl-Qa-fluoroi- NOCllTdIll lezshmamz 6a,1fia-dimethyl-tla-fiuoro-1,4-

preguene-17a,21-diol-3,11,20- ATCC-6855. preg1adiene-17a,21-diol-3,ll,20- trione. trione.

4. 6a,16a-dirnethy1-9m-fiuoro-4- Nocardiaformica NRRL- 6a,16a-dimethyl-9a-fluoro-1,4-

pregnene-116,l7a,21-triol-3,20- 247D. geginadiene-1]fl,l7a,21-triol-3- dione. one.

5 6a,16a-dimethyl-4-pregnene-11B, Mycobacterium phlei 6a,lfia-dimethyl-l,4-pregnadienel7a,2l-triol-3,20-dione. ATCC-12, 298. 1l5,17a,2l-tri0l-3,20-d1one.

6 6a,16a-dimethyl-4-pregneue-l7a, Mycobacterium Zactz'cola 6a,lfia-dimcthyl-l,i-pregnadiene- 21-dicl-3,11,20-t1ione. ATCG-2l, 297. 170:,21-di0l-3,11,20-trione.

and inoculated with about 2.5 to 5 ml. of a culture of Mycobacterium smegmatis (NRRL 13-1667) microorganisms, and the inoculated culture is then incubated at a temperature of 28 C., with agitation, for a 24-hour period. To the resulting culture is added a solution containing 10 mg. of 6a,l6a-dimethyl-9afiuoro-4-pregnenel lfi,l7a,2l-triol-3,20-dione dissolved in 0.2. ml. of dimethylformamide. The culture containing the steroid compound is incubated, with agitation, for an additional period of about 24 hours at 28 C.

The fermentation broth is extracted with three 50 ml.-

portions of ethyl acetate, and the ethyl acetate extracts are combined and evaporated in vacuo to a volume of about 5 ml. The concentrated solution is then used to prepare streak-paper chromatograms which are developed utilizing formamide as the stationary liquid phase and 50% benzene-50% chloroform as the mobile liquid phase. 'Ivvo bands are secured, one of which corresponds to the more mobile component, the 606,1611-di11'16thY1-90L- fluoro-4-pregnene-ll 8,17a,21-triol-3,20-dione starting material, and the other corresponds to the less mobile component, the A -dehydro derivative. The paper chromatog'ram is dried, and the latter band is cut off and extracted with methanol. The material extracted with methanol is again subjected to streak-paper chromatography, using paper which has been extracted for 48 hours with metha- 1101, and employing the chloroform-formamide system previously employed. The resulting chromatogram shows only a trace band corresponding to the starting material with the major band that corresponding to the less mobile component, the A -dehydro derivative. The paper chromatogram is thoroughly dried, and the band corresponding to the less mobile component is cut ofi'and extracted with methanol. The methanol extract is evaporated to dryness in vacuo to give 60L,16oc-di1'n6thYl-9a-flHOIO-l,4- pregnadiene-l 1 3, l7a,21-triol-3 ,ZO-dione. The 6a,l6 x dimethyl 9u-fluoro-1, 4-pregnadiene-11,8, 17a,2l-triol-3,20-dione is reacted with excess acetic anhydride in pyridine to give the 21-acetyl derivative which is purified by recrystallization from benzene-petroleum ether to give substantially pure 61x,16oc-difl16thYl-9a-fill0l0- 1,4-pregnadiene-l 1,3, 170;,21-triol-3,2i)-dione ZI-acetate.

Example 11 About 400 mg. of 3-ethylenedioxy-17a,20,'20,2l-bismethylenedioxy-l6a-methyl-S,6-epoxy-pregnane-1l-one is dissolved in about 100 ml. of benzene, and the solution purged with nitrogen. To the resulting solution is added about 3.7 ml. of a 3-molar solution of methyl magnesium bromide, and the resulting mixture is stirred at a temperature of about 70 C. in a nitrogen atmosphere for a period of about 5 hours. The reaction solution is cooled to about 0-5 C., and approximately 9 g. of ammonium chloride in ml. of Water is added over a 20-minute period. The layers are separated, and the aqueous layer extracted with a small portion of benzene. The benzene extracts are combined, washed with three 25-ml. portions of water, dried over magnesium sulfate, and evaporated to dryness in vacuo to give 3-ethylenedioxy-l7a,2(),20,21- bismethylenedioxy 6fi,16a dimethyl allopregnane 5a ol-ll-one.

Approximately mg. of S-ethylenedioxy-17a,2(),20, 21 bismethylenedioxy 65,160: dimethyl allopregnane-Su-ol-ll-one is dissolved in about 5 ml. of methanol, and the solution is purged with nitrogen. To this solution is then added 0.4 ml. of an 8% aqueous sulfuric acid solution, and the resulting mixture is heated under reflux for a period of about 35 minutes in a nitrogen atmosphere. At the end of this reflux period, the reaction solution is cooled to 05 C., and to the solution is slowly added, over a lO-minute period, a solution of approximately 0.4 g. of sodium carbonate in 12 ml. of water. The resulting slurry is cooled, and the precipitated material is recovered and dried to give l7cx,20,20,2l-bismethylenedioxy 6fl,16a dimethyl ailopregnane 3,11 dione-5u-ol. 7

About 100 mg. of 17a,20,20,2l-bismethylenedioxy-6fi, 16a-dimethyl-allopregnane-S,1l-dione-5a-ol is mixed with about 35 cc. of an 0.25% solution of potassium hydroxide in methanol; the resulting mixture is heated under reflux for a period of approximately 2 hours under a nitrogen atmosphere. The reaction mixture is then diluted with Water, and the aqueous mixture extracted with ethyl acetate. The ethyl acetate extracts are dried, evaporated to dryness, and the residual material issubjected to chromatographic purification using about 10 g. of washed alumina, utilizing benzene and mixtures of benzene and ether, as the eluting solvents. The product obtained by evaporation of the ether-rich fractions is recrystallized from mixtures of ethyl acetate and ether to give substantially pure 17a,20,20,2l bismethylenedioxy-6,8,16a-dimethyl-4- pregnene-3,11-dione.

About 0.1 gram of 17u,20,20,2l-bismethylenedioxy-Gfl, 16a-dimethyl-4-pregnene-3,1l-dione is suspended in 18 cc. of 50% aqueous acetic acid, and the suspension is carefully purged with nitrogen, following which the suspension is heated on a steam bath under nitrogen for a period of about 8 hours and then allowed to stand at room temperature for an additional period of 15 hours. The reaction solution is evaporated to dryness in vacuo to give 6fi,16u dimethyl-4-pregnene-17a,21-diol-3,11,20-trione. This material is dissolved in 20 cc. of chloroform, and the chloroform solution evaporated to dryness in vacuo. To the residual dry material is added 1 cc. of pyridine and 1 cc. of acetic anhydride, and the resulting mixture is heated on a steam bath for a period of about 15 minutes. The reaction solution is cooled to about O-S" C. and poured into ice water. The aqueous suspension is extracted with chloroform, and the chloroform extract is washed with aqueous sodium bicarbonate solution, dried over anhydrous sodium sulfate, and evaporated to dryness in vacuo. The residual material, dissolved in 30 cc. benzene, is adsorbed on grams of acid-Washed alumina; the adsorbate is washed with 50 cc. ether. The adsorbate is then eluted with 250 cc. of a 5:5 mixture of ether-chloroform, and then with 50 cc. of a 3:7 mixture of ether-chloroform. The eluates are evaporated in vacuo, and the residual material is crystallized from methanol to give 613,16u-dimethyl-4-pregnene-17a,21-diol-3,11, 20-trione 21-acetate.

To a solution of 100 mg. of 6B,16a-dimethyl-4-pregnene-l7e,21-diol-3,11,20-trione 21-acetate, in 6 ml. tbutanol, and 10.01 ml. of glacial acetic acid is added 70 mg. of selenium dioxide. The mixture is heated under reflux in a nitrogen atmosphere for a period of about 6 hours. The reaction solution is filtered and evaporated in vacuo. The residual material is dissolved in benzene, and the benzene solution is washed With an aqueous sodium bicarbonate solution and dried over anhydrous sodium sulfate. The benzene solution is stirred with mercury overnight, then centrifuged, and the clear benzene solution is evaporated in vacuo. The residual material thus obtained is separated using partition chromatography on diatomaceous silica (Supercel) to give 65,16- dimethyl-l,4-pregnadiene-17a,21-diol-3,l1,20 trione 21- acetate.

Example 12 The procedure of Example 2 is repeated but using, in place of the 17a,20,20,2l-bismethylenedioxy-6a,16ix-dimethyl-4-pregnene-3,1l-dione starting material therein employed, 100 mg. of 17o,20,20,2l-bismethylenedioxy- 6B,16a-dimethyl-4-pregnene-3,1l-dione, prepared as described in Example 11 hereinabove. In accordance with this procedure there is obtained, via the intermediate formation of 17a,20,20,2l-bismethylenedioxy-6B,16a-dimethyl 1,4 pregnadiene 3,11 dione, as final products 65,160: dimethyl 1,4 pregnadiene-17u,21-diol-3,11,20- trione and 6,8,16m-dimethyl-1,4-pregnadiene-l7a,21-diol- 3,11,20-trione Zl-acetate.

Example 13 About 5 g. of B-ethylenedioxy-17u,20,20,2l-bismethylenedioxy-6l3,16a-dimethyl-allopregnane 11 one 50c 01, Which can be prepared as described in Example 11 hereinabove, is dissolved in about 85 cc. of benzene. This solution is added to a stirred suspension containing 5 grams of lithium aluminum hydride in one liter of anhydrous ether, and the resulting suspension is heated under reflux for a period of about 4 hours. The reaction mixture is allowed to cool to about room temperature, and approximately 33 cc. of ethyl acetate is added to destroy excess lithium aluminum hydride. The resulting mixture is admixed With about cc. of water, the ethereal layer is decanted, and the aqueous layer, diluted with an additional 200 cc. of water, is extracted with chloroform. The ether and chloroform layers are combined, and the combined organic layers are dried over anhydrous sodium sulfate and evaporated to dryness in vacuo. The residual material, dissolved in about 250 cc. of benzene, is adsorbed on about 150 grams of acidwashed alumina, and the resulting adsorbate is eluted utilizing mixtures of petroleum ether-ether. This eluate is evaporated in vacuo, and the residual material is purified by crystallization from ether to give 3-ethylenedioxy- 17a,20,20,21 bismethylenedioxy 65,1604 dimethylal1opregnane-5a,1lfi-diol.

Approximately 100 mg. of 3-ethylenedioxy-17a,20,20, 21 bismethylenedioxy 65,160: dimethyl allopregnane- 50,1lfl-di0l is dissolved in about 5 ml. of methanol, and the solution is purged with nitrogen. To this solution is then added 0.4 ml. of an 8% aqueous sulfuric acidl solution, and the resulting mixture is heated under reflux for a period of about 35 minutes in a nitrogen atmosphere. At the end of this reflux period, the reaction solution is cooled to 05 C., and to the solution is slowly added, over a 10-minute period, a solution of approximately 0.4 g. of sodium carbonate in 12 ml. of Water. The resulting slurry is cooled, and the precipitated material is recovered and dried to give 17u,20,20, 2l-bismethylenedioxy 65,16 dimethyl allopregnane- 5 0 .,11B-di0l-3-0116.

About 100 mg. of 17o,20,20,2l-bismethylenedioxy- 6/3,l6oc dimethyl allopregnane 5 diol 3 one is mixed with about 35 cc. of an 0.25% solution of potassium hydroxide in methanol; the resulting mixture is heated under reflux for a period of approximately 2 hours under a nitrogen atmosphere. The reaction mixture is then diluted with water, and the aqueous mixture extracted With ethyl acetate. The ethyl acetate extracts are dried, evaporated to dryness, and the residual material is subjected to chromatographic purification using about 10 g. of washed alumina, utilizing benzene, and mixtures of benzene and ether, as the eluting solvents. The product obtained by evaporation of the etherrich fractions is recrystallized from mixtures of ethyl acetate and ether to give substantially pure 17a,20,20,2lbismethylenedioxy 65,16or dimethyl 4 pregncne- 11B-ol-3-one.

About 0.1 gram of 17u,20,20,21bismethylenedioxy 6,8,1606 dimethyl 4 pregnene 11/3 ol 3 one is suspended in 18 cc. of 50% aqueous acetic acid, and the suspension is carefully purged with nitrogen, following which the suspension is heated on a steam bath under nitrogen for a period of about 8 hours and then allowed to stand at room temperature for an additional period of 15 hours. The reaction solution is evaporated to dryness in vacuo to give 6,8,16a-dimethyl-4-pregnene- 11B,l7ot,21-triol-3,20-dione. This material is dissolved in 20 cc. of chloroform, and the chloroform solution evaporated to dryness in vacuo. To the residual dry material is added 1 cc. of pyridine and 1 cc. of acetic anhydride, and the resulting mixture is heated on a steam bath for a period of about 15 minutes. The reaction solution is cooled to about 05 C. and poured into ice Water. The aqueous suspension is extracted with chloroform, and the chloroform extract is washed with aqueous sodium bicarbonate solution, dried over anhydrous sodium sulfate, and evaporated to dryness in vacuo. The residual material, dissolved in 30 cc. benzene, is adsorbed on 5 grams of acid-washed alumina; the adsorbate is washed With 50 cc. ether. The adsorbate is then eluted with 250 cc. of a 5:5 mixture of ether-chloro form, and then with 50 cc. of a 3:7 mixture of etherchloroform. The eluates are evaporated in vacuo, and the residual material is crystallized from methanol to give 65,160 dimethyl 4 -pregnene 11,13,17rz,2l trio]- 3,20-dione Zl-acetate.

To a solution of 100 mg. of 65,16a-dimethyl-4-pregnene-l113,l7a,2l-triol-3,20-dione 21-acetate, in 6 ml. tbutanol, and 0.01 ml. of glacial acetic acid, is added 70 mg. of selenium dioxide. The mixture is heated under reflux in a nitrogen atmosphere for a period of about 6 hours. The reaction solution is filtered and evaporated in vacuo. The residual material is dissolved in benzene, and the benzene solution is washed with an aqueous sodium bicarbonate solution and dried over anhydrous sodium sulfate. The benzene solution is stirred with mercury overnight, then centrifuged, and the clear benzene solution is evaporated in vacuo. The residual material thus obtained is separated using partition chromatog raphy on diatomaceous silica (Supercel) to give 65,160;- dimethyl-1,4-pregnadiene-11fi,17u,21-triol-3,20-dione 21- acetate.

Example 14 To a cooled solution of 436 mg. of 6fi,l6a-dimethyl-4- pregnene-l1,8,l7a,2l-triol-3,20-dione 21-acetate (which can be prepared as described in Example 13 hereinabove) in 2.5 cc. dimethyl formamide and 2.0 ml. of dry pyridine is added 1.0 ml. of methane sulfonyl chloride, While maintaining the temperature below C. The resulting mixture is allowed to warm to room tempera ture, at which point a precipitate appears; the resulting mixture is then heated to a temperature of about 70- 100 C. for a period of about 10 minutes. About 15 ml. of water is added slowly to the reaction mixture, with stirring, and the aqueous mixture is extracted with ethyl acetate. The combined ethyl acetate extracts are washed With water, then with dilute aqueous hydrochloric acid solution, again with water, and then with a dilute aqueous sodium bicarbonate solution. The Washed ethyl acetate solution is dried, and the solvent is evaporated in vacuo; the residual material is triturated with ether, and the crystalline material is recrystallized to give 6B,l6oz-dimethyl-4,901)-pregnadiene-17a-2l-diol-3,20 dione 21- acetate. -A suspension of 330 mg. of 6fi,16u-dimethyl-4,- 9(l1)-pregnadiene-l7a,2l-diol-3,20-dione 2l-acetate and 1.8 g. of N-bromo-succinimide in a mixture of -0 ml. of dioxane and ml. of water is cooled to 10 C. Then with stirring, 10 ml. of cold 1.0 N aqueous perchloric acid is added. The temperature of the reaction mixture is allowed to rise to C. and is maintained at this point for about two-and-one-half hours during which time the solid material slowly dissolves. The resulting yellow solution is treated with 1.0 ml. of allyl alcohol to discharge the color and the remaining N-hromo-succinimide, and the remaining solution is evaporated to a small volume in vacuo. The concentrated solution is diluted with water, and the aqueous mixture is extracted with ethyl acetate. The ethyl acetate extracts are washed, dried and evaporated to dryness, and the residual material is crystallized from ethyl acetate-ether to give 941- bromo-6f3,l6m-dimethyl-4-pregnenel1 8,17a,21-triol 3, ZO-dione Zl-acetate.

A solution of 210 mg. of 9u-bromo-6fi,l6a-diniethyl-4- pregnene-llfl,l7oc,2l-triol-3,20-dione 21-acetate and 240 mg. of potassium acetate in 10 ml. of absolute ethanol is heated under reflux for two hours. The reaction mixture is cooled to room temperature, evaporated in vacuo to a small volume, and diluted with water. The concentrated aqueous mixture is extracted with three portions of ethyl acetate, and the combined ethyl acetate extracts are washed with water, dried, and evaporated to dryness in vacuo. The residual material is crystallized from ethyl acetate-ether to give 6,6,16a-dimethyl-9,l1- epoxy-4-pregnene-17a,21-dio1-3,20-dione 21-acetate.

To a solution of 83 mg. of anhydrous hydrogen fluoride in 4.7 of ice-cold alcohol-free chloroform is added an ice-cold solution of 416 mg. of 6fl,l6a-dim ethyl-9,11- epoxy-4-pregnene-l7a,21-diol-3,20-dione 2l-acetate. The solution is mixed thoroughly and maintained at 0 C. for two hours, at the end of which time 13.0 ml. of icecold 20% aqueous sodium acetate is added, and the resulting mixture agitated vigorously. The layers are separated, and the chloroform layer is washed with water,

until free of acid, dried, and the chloroform evaporated in vacuo. The residual material is crystallized from acetone-petroleum ether to give 65,16a-dimethyl-9afluoro-4-pregnene-l113,l7a,21-triol-3,20-dione 2l-acetate. Fifty milligrams of 65,l6ez-dimethyl-9a-fluoro-4-pregnene- 11/3,17a,2l-triol-3,20-dione 2l-acetate is dissolved in a mixture of 1.0 cc. of benzene and 1.0 cc. of l N methanolic potassium hydroxide, and the solution is allowed to stand at room temperature for a period of about 10 minutes. The solution is then acidified with acetic acid, the benzene is evaporated in vacuo, and the residual material is purified by crystallization to give 6fi,16oc-dimethyl-9a-fluoro-4-pregnene-1 1,8, l7u,2l-triol-3,20-dione.

To a solution of mg. of 65-l6wdimethyl 9oc-flll01'0- 4-pregnene-11fl,17u,2l-triol-3,20-dione ZI-acetate, in 6 ml. t-butanol, and 0.01 ml. of glacial acetic acid is added 70 mg. of selenium dioxide. The mixture is heated under reflux in a nitrogen atmosphere for a period of about 60 hours. The reaction solution is filtered and evaporated in vacuo. The residual material is dissolved in benzene, and the benzene solution is washed with an aqueous sodium bicarbonate solution and dried over anhydrous sodium sulfate. The benzene solution is stirred with mercury overnight, then centrifuged, and the clear benzene solution is evaporated in vacuo. The residual material thus obtained is separated, using partition chromatoggraphy on diatomaceous silica (Supercel), to give 613,1602, dimethyl-9ot-fluoro-1,4-pregnadiene-l l fl,17u,21-t.riol 3,20- dione ZI-acetate. This material is hydrolyzed using 1 N methanolic potassium hydroxide in benzene as described above for the hydrolysis of 63,l6a-dimethyl-9a-fluoro-4- pregnene-l1 B,17a,21-triol-3,20-dione ZI-acetate to give 6B,16a-dimethyl-9m-fluoro-L4 pregnadiene llfi,17a,21- triol-3,20-dione.

Example 15 The procedure of Example 6 is repeated but using, in place of the 6a,l6m-dimethyl-9a-fluoro-4-pregnene-11B,- l7a,2l-triol-3,20-dione 2l-acetate employed in that example, 400 mg. of 6,8,16wdimethyl-9a-fluoro-4-pregnene 11,8,17a,21-t1l0l3,20-dl0116 ZI-acetate, which can be prepared as described in Example 14 hereinabove. In accordance with this procedure there are obtained as products 6/3,l6a-dimethyl-9a-fluoro-4-pregnene-l7a,21- diol-3,1 1,20-trione 2l-acetate; 6fl,l6a-dimethyl-9u-fluoro 4-pregnene-17u,21-diol-3,l1,20-trione; 6 8,16a-dimethyl- 9a-fluoro-1,4-pregnadiene-l7a,21-di0l-3,l1,20-trione 21- acetate and 65,16wdimethyl-9a-fluoro-1,4-pregnadienel7tx,2l-diol-3,11,20-trione, respectively.

Example 16 10 grams of 16(3-methyl-4-pregnene-17a,2l-dio1-3,ll,- 20-trione are dissolved in 400 cc. of chloroform. To this solution is added a mixture of 100 cc. of concentrated aqueous hydrochloric acid and 100 cc. of 37% aqueous formaldehyde solution. The resulting mixture is stirred for a period of approximately 3 days at room temperature. The chloroform layer is separated, washed with sodium bicarbonate solution until neutral, then with water, dried over anhydrous sodium sulfate, and evaporated to dryness in vacuo. The residual crude material is purified by crystallization from a mixture of methylene chloride and methanol to give l7,1,20,20,21-bismethylenedioxy-166- methyl-4-pregnene-3 ,1 l-dione.

10grams of l7oz,20,20,21-bismethylenedioxy-l6 3-meth yl-4-pregnene-3,ll dione are dissolved in 500 cc. of benzene, to the solution is added 25 cc. ethylene glycol and 1 gram ofp-toluenesulfonic acid, and the resulting mix- 33 ture is heated under reflux for a period of approximately 15 hours. The reaction solution is cooled, washed with aqueous sodium bicarbonate solution, dried over anhydrous sodium sulfate, and evaporated to dryness in vacuo.

The residual crude material is adsorbed from its solution in benzene on 250 g. of acid-washed alumina, and the resulting adsorbate is eluted utilizing mixtures of etherpetroleum ether to give 3-ethylenedioxy-17a,20,20,21-bismethylenedioxy- 1 6B-methyl-5 -pregnene-1 l-one.

A mixture of 10 grams of 3-ethylenedioxy-17a,20,20,- 21-bismethylenedioxy-16,8-methyl-5-pregnene-1l-one, 4.3 g. perbenzoic acid and 230 cc. benzene is allowed to stand at room temperature for a period of about 2 days. A portion of the epoxide product, which precipitates, is separated from the reaction solution by decantation, and the decanted solution is washed with saturated aqueous sodium bicarbonate solution and then dried over anhydrous sodium sulfate. The dried solution is then evaporated in vacuo, and the residual epoxide product is combined with the epoxide product originally separated from the reaction mixture to give 3:ethylenedioxy-l7a,20,20,2l-bismethylenedioxy-l 6,8-methyl-5 ,6-epoxy-pregnane-l l-one.

The latter product is dissolved in 200 cc. of substantially anhydrous formic acid and the resulting solution is allowed to stand at room temperature for a period of about 2%. hours. The reaction solution is poured into water, and the aqueous mixture is extracted with chloroform. The chloroform solution is washed with saturated aqueous sodium bicarbonate solution, dried over anhydrous sodium sulfate, and evaporated in vacuo to give a mixture of 17m,20,20,2l-bismethylenedioxy-165-methyl- 6-formyloxypregnane-S-ol-3,1l-dione and l7a,20,20,21- bismethylenedioxy 1615 methyl 5 formyloxy pregnane-6-ol-3,11-dione.

This mixture of S-formyloxy and 6-formyloxy derivatives is dissolved in about 850 cc. of methanoL'to this solution is added a solution containing about 17 grams of potassium hydroxide in 80 cc. of water, and the resulting mixture is heated under reflux in a nitrogen atmosphere for a period of approximately /2 hour. The reaction solution is cooled, neutralized with about 23 cc. of acetic acid and evaporated in vacuo to a small volume. The concentrated solution is poured into water and the aqueous mixture is extracted with chloroform. :The chloroform extract is washed with aqueous sodium bicarbonate solution, dried over anhydrous sodium sulfate, and evaporated to dryness in vacuo. The residual material is crystallized from methanol to give l7u,20,20,2l-bismethylenedioxy- 1613-methyl-allopregnane-3,6,1l-trione.

A solution containing about 5 grams of 1704,2020,- 21 bismethylenedioxy 16,8 methyl allopregnane 3,- 6,11-trione and 0.1 g. of p-toluenesulfonic acid in 120 cc. butanone dioxolane is heated under reflux for a period of about 10 minutes. The reaction solution is cooled to about 5 C., diluted with chloroform, and the resulting solution is washed with aqueous sodium bicarbonate solution and dried over anhydrous sodium sulfate. The resulting dried solution is evaporated to dryness in vacuo, and the residual material is slurried with ether, recovered by filtration and then dried to give 3-ethylenedioxy-l7a; 20,20,21 bismethylenedioxy 1618 methyl allopregnane-6,11-dione.

A solution containing about grams of 3-ethylenedioxy l7a,20,20,21 bismethylenedioxy 16B methylallopregnane-6,l1-dione in 70 cc. of benzene is added, with stirring over a period of approximately minutes, to an ethereal solution containing the methyl magnesium iodide, prepared from Sec. of methyl iodide and 0.5 gram of magnesium turnings, dissolved in 50 cc. of ether. The reaction mixture is allowed to stir for an additional /2 hour period, and the resulting solution is decomposed with 70 cc. of water. About 170 cc. of benzene is added to the aqueous mixture and the layers are separated. The aqueous layer is extracted with two 100' cc.-portions of chloroform, and the organic layers are combined, dried 34 over anhydrous sodium sulfate, and evaporated to dryness in vacuo to give 3-ethylenedioxy-17ot,20,20,2l-bismethylenedioxy-6u,16,8-dimethyl-allopregnane-6fi-ol-l l-one.

A solution containing 5.5 cc. of freshly distilled thionyl chloride in 26 cc. of cold anhydrous pyridine is added dropwise, with stirring, to a solution of 5.0 g. of 3-ethylenedioxy l7u,20,20,21 bismethylenedioxy 60L, 1 6B dimethylallopregnane-65-ol-1l-one in 32 cc. of anhydrous pyridine, while maintaining the temperature of the reaction mixture at approximately 40 C. The reaction solution is stirred for an additional 30-minute period following the addition of the thionyl chloride reagent, and the reaction mixture is then cooled to about 05 C. and poured into 180 cc. ice water. The aqueous mixture is extracted with chloroform, and the chloroform extract is neutralized, dried over anhydrous sodium sulfate, and evaporated to dryness in vacuo. The residual material, dissolved in benzene, is chromatographed on grams of acid-Washed alumina; elution of the resulting adsorbate with mixtures of petroleum ether and ether gives 3-ethylenedioxy-17a, 20,20,21 bismethylenedioxy 6,165 dimethyl 5 pregnene-l l-one.

A solution containing about 5 grams of 3-ethylenedioxy- 17oz-20,20,21 bisrnethylenedioxy 6,16B-dimethyl-5-pregnene-ll-one, 500 cc. of anhydrous acetone and about 0.5 g. of p-toluenesulfonic acid monohydrate is allowed to stand for a period of about 15 hours. The reaction solution is poured into water, and the aqueous mixture is extracted with chloroform. The chloroform solution is washed with aqueous sodium bicarbonate solution, dried over anhydrous sodium sulfate, and the chloroform solution is evaporated to dryness in vacuo. The residual material is crystallized from benzene to give L,20,20,2lbismethylenedioxy 60,16B dimethyl 4 pregnene-3,l1 dione.

About 0.1 gram of 17ot,20,20,2l-bismethylenedioxy-6a, 1GB-dimethyl-4-pregnene-3,1l-dione is suspended in 18 cc. of 50% aqueous acetic acid, and the suspension is carefully purged with nitrogen, following which the suspension is heated on a steam bath under nitrogen for a period of about 8 hours and then allowed to stand at room temperature for an additional period of 15 hours. The reaction solution is evaporated to dryness in vacuo to give 6a,l6/3-dimethyl-4-pregnene-17a,21-diol-3,11,20- trione. This material is dissolved in 20 cc. of chloroform, and the chloroform solution evaporated to dryness in vacuo. To the residual dry material is added 1 cc. of pyridine and 1 cc. of acetic anhydride, and the resulting mixture is heated on a steam bath for a period of about 15 minutes. The reaction solution is cooled to about 05 C. and poured into ice water. The aqueous suspension is extracted with chloroform, and the chloroform extract is Washed with aqueous sodium bicarbonate solution, dried over anhydrous sodium sulfate, and evaporated to dryness in vacuo, The residual material, dissolved in 30 cc. benzene, is adsorbed on 5 grams of acid-washed alumina; the adsorbate is Washed with 50 cc. ether. The adsor-bate is then eluted with 250 cc. of a 5:5 mixture of ether-chloroform, and then with 50 cc. of a 3:7 mixture of ether-chloroform. The eluates are evaporated in vacuo, and the residual material is crystallized from methanol to give 6a,l6/3-dimethyl- 4-pregnene-17a,21-diol-3,11,20-trione 21-acetate.

To a solution of 100 mg. of'6u,lofi-dimethyli-pregnene- 17a,21-diol-3,1-l,20-trione 21-acetate, in 6 -ml. t-butanol, and 0.01 ml. of glacial acetic acid is added 70 mg. of selenium dioxide. The mixture is heated under reflux in a nitrogen atmosphere for a period of about 6 hours. The reaction solution is filtered and evaporated in vacuo. The residual material is dissolved in benzene, and the benzene solution is washed with an aqueous sodium bicarbonate solution and dried over anhydrous sodium sulfate. The benzene solution is stirred with mercury overnight, then centrifuged, and the clear benzene solution is evaporated in vacuo. The residual material thus 35 obtained is separated using partition chromatography on 'diatomaceous silica (Supercel) to give 6a,l6fi-dimethyl- 1,4-pregnadiene-l7a,2l-diol-3,11,20-trione 21-acetate.

Example 17 About 400 mg. of 3-ethylenedioxy-17a,20,20,21-bismethylenedioxy-l6fi-methyl-5,6-epoxy-pregnane-1l-one is dissolved in about 100 ml. of benzene, and the solution purged with nitrogen. To the resulting solution is added about 3.7 ml. of a 3-molar solution of methyl magnesium bromide, and the resulting mixture is stirred at a temperature of about 70 C. in a nitrogen atmosphere for a period of about 5 hours. The reaction solution is cooled to about -5 C., and approximately 9 g. of ammonium chloride in 90 ml. of water is added over a 20-minute period. The layers are separated, and the aqueous layer extracted with a small portion of benzene. The benzene extracts are combined, washed with three 25-ml. portions of water, dried over magnesium sulfate, and evaporated to dryness in vacuo to give 3-ethylenedioxyl7et,20,20,2l bismethylenedioxy 65,16,8- dimethylallopregnane-5a-ol-1 l-one.

Approximately 100 mg. of 3-ethylenedioXy-17a,20,20, 21 bismethylenedioxy 6 8,165 dimethyl allopregnane- 5a-ol-11-one is dissolved in about 5 ml. of methanol, and the solution is purged with nitrogen. To this solution is then added 0.4 ml. of an 8% aqueous sulfuric acid solution, and the resulting mixture is heated under reflux for a period of about 35 minutes in a nitrogen atmosphere. At the end of this reflux period, the reaction solution is cooled to 05 C., and to the solution is slowly added, over a 10-minute period, a solution of approximately 0.4 g. of sodium carbonate in 12 ml. of water. The

resulting slurry is cooled, and the precipitated material is recovered and dried to give l7a,20,20,2l-bismethylenedioxy-6l5,16;3-dimethyl-allo-pregnane-3 .1 l-dione-5 u-ol.

About 100 mg. of l7u,20,20,2l-bismethylenedioxy- 65,165-dimethyl-allophegnane-3,l1-dione-5a-ol is mixed with about 35 cc. of an 0.25% solution of potassium hydroxide in methanol; the resulting mixture is heated under reflux for a period of approximately 2 hours under a nitrogen atmosphere. The reaction mixture is then diluted with water, and the aqueous mixture extracted with ethyl acetate. The ethyl acetate extracts are dried, evaporated to dryness, and the residual material is subjected to chromatographic purification using about 10 g. of washed alumina, utilizing benzene, and mixtures of benzene and ether, as the eluting solvents. The product obtained by evaporation of the ether-rich fractions is recrystallized from mixtures of ethyl acetate and ether to give substantially pure 170:,20,20,21 bismethylenedioxy 63, 16J8-dimethyl-4-pregnene3 ,1 l-dione.

About 0.1 gram of 17a,20,20,21-bismethylenedioxy- 6,8,16,8-dimethyl-4-pregnene-3,1l-dione is suspended in 18 cc. of 50% aqueous acetic acid, and the suspension is carefully purged with nitrogen, following which the suspension is heated on a steam bath under nitrogen for a period of about 8 hours and then allowed to stand at room temperature for an additional period of hours. The reaction solution is evaporated to dryness in vacuo to give 6B,16,8-dimethyl-4-pregnene-170:,21-di0l-3,11,20

' trione. This material is dissolved in 20 cc. of chloroform,

and the chloroform solution evaporated to dryness in vacuo. To the residual dry material is added 1 cc. of pyridine and 1 cc. of acetic anhydride, and the resulting mixture is heated on a steam bath for a period of about 15 minutes. The reaction solution is cooled to about 0-5 C. and poured into ice water. The aqueous suspension is extracted with chloroform, and the chloroform extract is washed 'with aqueous sodium bicarbonate solution, dried over anhydrous sodium sulfate, and evaporated to dryness in vacuo. The residual material, dissolved in 30 cc. benzene, is adsorbed on 5 grams of acid-washed alumina; the adsorb'ate is washed with 50 cc. ether. The adsorbate is then eluted with 250 cc. of a 5:5 mixture of 36 ether-chloroform, and then with 50 cc. of a 3:7 mixture of ether-chloroform. The eluates are evaporated in vacuo, and the residual material is crystallized from methanol to give 66,166 dimethyl-4-pregnene-l7a,2l-diol-3,l1,20-trione ZI-acetate.

To a solution of 100 mg. of 6,6,16fi-dimethyl-4-pregnene-17a,21-diol-3,11,20-trione 2l-acetate, in 6 ml. t-butanol, and 0.01 ml. of glacial acetic acid is added 70 mg. of selenium dioxide. The mixture is heated under reflux in a nitrogen atmosphere for a period of about 6 hours. The reaction solution is filtered and evaporated in vacuo. The residual material is dissolved in benzene, and the benzene solution is washed with an aqueous sodium bicarbonate solution and dried over anhydrous sodium sulfate. The benzene solution is stirred with mercury overnight, then centrifuged, and the clear benzene solution is evaporated in vacuo. The residual material thus obtained is separated using partition chromatography on diatomaceous silica (Supercel) to give 6p,l6e-dimethyl-l,4-pregnadiene-l7oc,2l-diol-3, 1 1,20-trione 2l-acetate.

Example 18 The procedure of Example 2 is repeated but using, in place of the l7a,20,20,2l-bismethylenedioxy-6a,16x-dimethyl-4-pregnene-3,1l-dione starting material there employed, 100 mg. of 17a,20,20,2I-bismethylenedioXy-Ga, 16fi-dimethyl-4-pregnene-3,ll-dione, which can be prepared as described in Example 16 hereinabove. In accordance with this procedure there are obtained, via the intermediate formation of l7a,20,2 0,2l-bismethylenedioxy-6u,16t3-dimethyl-1,4-pregnadiene-3,ll-dione, the final products 6a,l6B-dimethyl 1,4 pregnadiene-17a,21-diol- 3,11,20-trione and 6a,16B-dimethy1-1,4-pregnadiene-l7a, 21-diol-3,l1,20-trione 21-acetate.

Example 19 The procedure of Example 2 is repeated but using, in place of the 17a,20,20,21-bismethylenedioxy-6a,l6a-dimethyl-4-pregnene-3,1l-dione starting material there employed, 100 mg. of 17a,20,20,2l-bismethylenedioxy- 6B,16B-dimethyl-4-pregnene-3,ll-dione, which can be prepared as described in Example 17 hereinabove. In accordance with this procedure there are obtained, via the intermediate formation of 17a,20,20,2l-bismethylenedioxy-6e,lGB-dimethyl-1,4-pregnadiene-3,ll-dione, the final products 6B,1GB-dimethyl-1,4-pregnadiene 17a,2l diol- 3,11,20 trione and 66,166 dimethyl 1,4-pregnadienel7ix,2l-diol-3,11,20-trione 21-acetate.

Example 20 About 5 grams of 3-ethylenedioxy-17 a,20,20,21-bismethylenedioxy-6,16,6-dimethyl-5-pregnene-l 1-one, which can be prepared as described in Example 16 hereinabove, is dissolved in about cc. of benzene. This solution is added to a stirred suspension containing 5 grams of lithium aluminum hydride in one liter of anhydrous ether, and the resulting suspension is heated under reflux for a period of about 4 hours. The reaction mixture is allowed to cool to about room temperature, and approximately 33 cc. .of ethyl acetate is added to destroy excess lithium aluminum hydride. The resulting mixture is admixed with about cc. of water, the ethereal layer is decanted, and the aqueous layer, diluted with an additional 200 cc. of water, is extracted with chloroform. The ether and chloroform layers are combined, and the combined organic layers are dried over anhydrous sodium sulfate and evaporated to dryness in vacuo. The residual material, dissolved in about 250 cc. of benzene, is adsorbed on about grams of acid-washed alumina, and the resulting adsorbate is eluted utilizing mixtures of petroleum etherether. This eluate is evaporated in vacuo, and the residual material is purified by crystallization from ether to give 3-ethylenedioxy 17a,20,20,2l bismethylenedioxy- 6, l6B-dimethyl-5-pregnene-l lB-ol.

A solution containing about 5 g. of 3-ethylenedioxyz,2(),20,21 bismethylenedioxy-6,16,8-dimethy1-5-pregnene-11,8-ol, 500 cc. of anhydrous acetone and about 0.5 g. of p-toluenesulfonic acid monohydrate is allowed to stand for a period of about 15 hours. The reaction solution is poured into water, and the aqueous mixture is extracted with chloroform. The chloroform solution is washed with aqueous sodium bicarbonate solution, dried over anhydrous sodium sulfate, and the chloroform solution is evaporated to dryness in vacuo. The residual material is crystallized from benzene to give 17a,20,2(),21- bismethylenedioxy 6a,l6fi-dimethyl-4-pregnene-11fl-0l-3- one.

About 0.1 gram of 17a,20,20,2l-bismethylenedioxy 6a,l6B-dimethyl-4-pregnene-11,8-ol-3-one is suspended in 18 cc. of 50% aqueous acetic acid, and the suspension is carefully purged with nitrogen, following which the suspension is heated on a steam bath under nitrogen for a period of about 8 hours and then allowed to stand at room temperature for an additional period of 15 hours. The reaction solution is evaporated to dryness in vacuo to give 6a,16,8 dimethyl 4-pregnene-l1fl,17u,21-tri0l-3,20- dione. This material is dissolved in 20 cc. of chloroform, and the chloroform solution evaporated to dryness in vacuo. To the residual dry material is added 1 cc. of pyridine and 1 cc. of acetic anhydride, and the resulting mixture is heated on a steam bath for a period of about 15 minutes. The reaction solution is cooled to about 5 C. and poured into ice water. The aqueou suspension is extracted with chloroform, and the chloroform extract is washed with aqueous sodium bicarbonate solution, dried over anhydrous sodium sulfate, and evaporated to dryness in vacuo. The residual material, dissolved in 30 cc. benzene, is adsorbed on 5 grams of acid-washed alumina; the adsorbate is washed with 50 cc. ether. The adsorbate is then eluted with 250 cc. of a 5:5 mixture of ether-chloroform, and then with 50 cc. of a 3:7 mixture of ether-chloroform. The eluates are evaporated in vacuo, and the residual material is crystallized from methanol to give 6u,16 3-dimethyl-4-pregnene-l1fi,17u,21- trio1-3,20-dione 21-acetate.

To a solution of 100 mg. of 6u,165-dimethyl-4- pregnene-l1B,l7oc,2l-triol-3,20-dione 21-acetate, in 6 ml. t-butanol, and 0.01 ml. of glacial acetic acid, is added 70 mg. of selenium dioxide. The mixtur is heated under reflux in a nitrogen atmosphere for a period of about 6 hours. The reaction solution is filtered and evaporated in vacuo. The residual material is dissolved in benzene, and the benzene solution is washed with an aqueous sodium bicarbonate solution and dried over anhydrous sodium sulfate. The benzene solution is stirred with mercury overnight then centrifuged and the clear benzen solution is evaporated in vacuo. The residual material thus obtained is separated using partition chromatography on diatomaceous silica (Supercel) to give 6a,16 8 dimethyl 1,4-pregnadiene-llB,l7ot,21triol-3,20- dione ZI-acetate.

Example 21 About 5 g. of 3-ethylenedioxy-17a,20,20,21-bismethylenedioxyl 6,8,16 8 dimethyl allopregnane-ll-one-Sa-ol, which can be prepared as described in Example 17 hereinabove, is dissolved in about 85 cc. of benzene. This solution is added to a stirred suspension containing 5 grams of lithium aluminum hydride in one liter of anhydrous ether, and the resulting suspension is heated under reflux for a period of about 4 hours. The reaction mixture is allowed to cool to about room temperature, and approximately 33 cc. of ethyl acetate is added to destroy excess lithium aluminum hydride. The resulting mixture is admixed with about 100 cc. of water, the ethereal layer is decanted, and the aqueous layer, diluted with an additional 200 cc. of water, is extracted with chloroform. The ether and chloroform layers are combined, and the combined organic layers are dried over anhydrous sodium sulfate and evaporated to dryness in vacuo. The residual material, dissolved in about 250 cc. of benzene, is adsorbed on about grams of acid-washed alumina, and the resulting adsorbate is eluted utilizing mixtures of petroleum ether-ether. This eluate is evaporated in vacuo, and the residual material is purified by crystallization from ether to give 3-ethylenedioxy-17a,20,20,21--bismethylenedioxy 6fi,l6,8 dimethyl-allopregnane-5or,11(3- diol.

Approximately 100 mg. of 3-ethylenedioxy-17a,20,20, 21 bismethylenedioxy 65,16fi dimethyl-allopregnane- 5a,11,8-diol is dissolved in about 5 ml. of methanol, and the solution is purged with nitrogen. To this solution is then added 0.4 ml. of an 8% aqueous sulfuric acid solution, and the resulting mixture is heated under reflux for a period of about 35 minutes in a nitrogen atmosphere. At the end of this reflux period, the reaction solution is cooled to 05 C., and to the solution is slowly added, over a 10-minute period, a solution of approximately 0.4 g. of sodium carbonate in 12 ml. of water. The resulting slurry is cooled, and the precipitated material is recovered and dried to give 17u,20,20,2l-bismethylenedioxy- 6,8,16,6-dimethyl-allopregnane-Sa,1 1,8-diol-3-one.

About 100 mg. of ,20,20,21-bismethylenedioxy-6/3- l6,8-dimethyl-allopregnane-5or,1 1 fl-diol-S -one is mixed with about 35 cc. of an 0.25% solution of potassium hydroxide in methanol, the resulting mixture is heated under reflux for a period of approximately 2 hours under a nitrogen atmosphere. The reaction mixture is then diluted with water, and the aqueous mixture extracted with ethyl acetate. The ethyl acetate extracts are dried, evaporated to dryness, and the residual material is subjected to chromatographic purification using about 10 g. of washed alumina, utilizing benzene, and mixtures of benzene and ether, as the eluting solvents. The product obtained by evaporation of the ether-rich fractions is recrystallized from mixtures of ethyl acetate and ether to give substantially pure 17a,20,20,21 bisrnethylenedioxy 65,165 dirnethyl-4-pregnene-l 1 B-ol-3-one.

About 0.1 gram of .17a.,20,20,2l-bismethylenedioxy- 6p,l6p-dimethyl-4-pregnene-l1,6-01-3-one is suspended in 18 cc. of 50% aqueous acetic acid, and the suspension is carefully purged with nitrogen, following which the suspension is heated on a steam bath under nitrogen for a period of about 8 hours and then allowed to stand at room temperature for an additional period of 15 hours. The reaction solution is evaporated to dryness in vacuo to give 65,165 dimethyl 4 pregnene 1lfl,17u,21 triol 3,20- dione. This material is dissolved in 20 cc. of chloroform, and she chloroform solution evaporated to dryness in vacuo. To the residual dry material is added '1 cc. of pyridine and 1 cc. of acetic anhydride, and the resulting mixture is heated on a steam bath for a period of about 15 minutes. The reaction solution is cooled to about 05 C. and poured into ice water. The aqueous suspension is extracted with chloroform, and the chloroform extract is washed with aqueous sodium bicarbonate solution, dried over anhydrous sodium sulfate, and evaporated to dryness in vacuo. The residual material, dissolved in 30 cc. benzene, is adsorbed on 5 grams of acid-washed alumina; the adsorbate is washed with 50 cc. ether. The adsorbate is then eluted with 250 cc. of a 5:5 mixture of ether-chloroform, and then with 50 cc. of a 3:7 mixture of ether-chloroform. The eluates are evaporated in vacuo, and the residual material is crystallized from methanol to give 66,16fi-dimethyl-4-pregnene-11fi,l7o,21-triol- 3,2.0-dione 21-acetate.

To a solution of 100 mg. of 6[3,16,8-dimethyl-4-preg nene-l1,8,l7e,21-diol-3,20-dione 21-acetate, in 6 ml. tbutanol, and 0.01 ml. of glacial acetic acid, is added 70 mg. of selenium dioxide. The mixture is heated under reflux in a nitrogen atmosphere for a period of about 6 hours. The reaction solution is filtered and evaporated in vacuo. The residual material is dissolved in benzene, and the benzene solution is washed with an aqueous sodium bicarbonate solution and dried over anhydrous sodium sulfate. The benzene solution is stirred with mer- 39 cury overnight then centrifuged and the clear benzene solution is evaporated in vacuo. The residual material thus obtained is separated using partition chromatography on diatomaceous silica (Supercel) to give 6B,165-dimethyll,4pregnadiene-l1/3,17 x,21-triol-3,20-dione Zl -acetate.

Example 22 The procedure of Example is repeated but using, in place of the 6a,l6a-dimethyl-4-pregnene-1lfi,17a,2l-triol- 3,20adione 21-acetate starting material there employed, about 435 mg. of 6a,lfl-dimethyll-pregnene-llfi,l7a,21- triol-3,20-dione 21-acetate, which can be prepared as described in Example hereinabove. In accordance with this procedure, there are obtained, via the intermediate formation of 6a,l6 3-dimethyl-4,9(1-1)-pregnadiene-17x, 21-dio1-3,20-dione 21-acetate; 9a.-bromo-6a,16fi-dimethyl- 4-pregnene-11B,17a,21-triol-3,20-dione Zl-acetate and 60:, 16,8 dimethyl 9,11 epoxy 4 pregnene 17a,21 diol- 3,20-dione ZI-acetate, as final products 6a,16p-dimethyl- 9a fluoro 4 pregnene llfi,l7cc,2l triol 3,20 dione 2 l-acetate; 6a, l6fi-dimethyl-9a-fluoro-4-pregnene-1 1,8, 17a, 21-triol-3,20-dione; 6a,16B-dimethy1-9a-fiuoro-1,4-pregnad-iene-l1,6,l7a.,2l-triol-3,20-dione 21-acetate and 60:,16/3- 'di-methyl 90c fiuoro 1,4 pregnadiene 11B,17a,21- triol-3,20-dione, respectively.

Example 23 The procedure of ExampleS is repeated, but using, in place of 6a,16a-dimethyl-4-pregnene-11fl,17u,21-triol-3, ZO-dione Zl-acctate starting material there employed, about 435 mg. of 6,8,16fi-dimethyl-4-pregnene-11153170 21- triol-3,20-dione 21-acetate, which can be prepared as described in Example 21 hereinabove. In accordance with this procedure, there are obtained, via the intermediate formation of 6fi,l6B-dimethyl-4,9(11)-pregnadiene-17a, 21-diol-3,20-dione ZI-acetate; 9tx-bromo-6,8,16fidimethyl 4-pregnene-llfi,l7a,21-tri0l-3,20-dione ZI-acetate and 6B, 166 dimethyl 9,11 --epoxy 4 pregnene 1701,21 diol- 3,20-dione 2l-acetate, as final products 65,165-dimethyl- 9a. fluoro 4 pregnene l1B,l7a,2l triol 3,20 dione 21 -acetate; 6p,16,8-dimethyl-9a-fluoro-4-pregnene- 1 113, 170:, 21-triol-3,20-dione; 66,16 3-dimethyl-9a-fluoro-1,4-pregnadiEIle-l1f3,1706,21et1i01-3,20-dl011 21-acetate and 65,165- dirnethyl 90c fluoro 1,4 pregnadiene 11B,17u,21- triol-3,20-dione, respectively.

Example 24 The procedure of Example 6 is repeated but using, in place of the 6a,l6a-dimethyl-9a-iiuoro-4-pregnene-11B, 17a,21-triol-3,20-dione 21-acetate starting material there employed, 400 mg. of 6a,16Bdimethyl-9a-fiuoro-4-pregnene-l115,17a,2l-triol-3,20-dione 21-acetate, which can be prepared as described in Example 22 hereinabove. In accordance with this procedure there are obtained as final products 6oz,16 8-dimethyl-9a-fiuoro 4 pregnene-17ot,2 ldiol-3,11,20-trione ZI-acetate; 6a,16;3-dimethyl-9a-fiuoro- 4-pregnene-17zx,21-diol-3,11,20-trione; 611,165 dimethyl- 9a-fluoro-l,4-pregnadiene-17a,2l-diol-3,11,20 trione 21- acetate and 6a,16;8-dimethyl 9a fiuoro-l,4-pregnadieue- 17a,21-diol-3,l1,20-trione, respectively.

Example 25 The procedure of Example 6 is repeated but using, in place of the 6a,l6a-dimethyl-9u-fluoro 4 pregnene-l 15, 17a,21-triol-3,20-dione 21-acetate starting material there employed, 400 mg. of 6,8,16,6-dimethyl-9a-fiuoro-4-pregnene-11,6,17a,2l-triol-3,20-dione ZI-acetate, which can be prepared as described in Example 23 hereinabove. In accordance with this procedure there are obtained as final products, 65,165 dimethyl-9ot-fluoro4-pregnene-17a,21- diol-3,11,20-trione 21-acetate; 66,16B-dirnethyl-9a-fluoro- 4-pregnene-17a,21-diol-3,l1,20-trione; 65,165 dimethyl- 9a-fluoro-1,4-pregnadiene-l7a,21-diol-3,l1,20 trione 21- acetate and 6;3,16;8dimethyl-9a-fiuoro 1,4 pregnadiene- 17 a,2ldi0l-3,1 1,20-trione, respectively.

acetate.

Example 26 p A suspension containing 300 mg. of 17a,21-dihydroxy- 6a,16a dimethyl 4 pregnene-3,l1,20-trione 2l-acetate, 700 mg. of chloranil, 8.3 ml. ofethyl acetate and 1.7 ml. of acetic acid is refluxed for 17 hours under nitrogen. The reaction mixture is diluted with 40 ml. of ethyl acetate and filtered. The filtrate is washed sequentially with two 20 ml. portions of ice cold 10% aqueous sodium bisulfite solution, three 20 ml. portions of ice cold 5% aqueous potassium hydroxide solution and finally washed to neutrality with several 20 m1. portions of ice water. The aqueous washes are re-extracted with 50 ml. of ethyl acetate and treated as above. The combined organic phase is dried over sodium sulfate and concentrated in vacuo. The crude material dissolved in benzene is chromatographed on 15 g. of acid-Washed alumina. Elution with chloroformzether 2:8 aflords l7ot,21-dihydroxy6, 16a-dimethyl 4,6 pregnadiene-3,11,20-trione 21-acetate. This product is then hydrolyzed by treatment with a solution of potassium bicarbonate in aqueous methanol to form 17a,21 dihydroxy-6,16ot-dimethyl-4,6-pregnadiene- 3,11,20-trione.

In accordance with the above procedure but starting with 17a,21 dihydroxy-6a,16,8-dimethyl-4-pregnene-3,11, ZO-trione 2l-acetate there are obtained 17u,21-dihydroxy- 6,l6fi-dimethyl-4,6-pregnadieue-3,11,-20-trione and its 21 acetate.

Example 27 A suspension containing 300 mg. of 11,8,17a,21-trihydroxy-6a,16adirnethyl-4-pregnene-3,20 dione ZI-acetate, 700 mg. of chloranil, 8.3 ml. of ethyl acetate and 1.7 ml. of acetic acid is refluxed for 17 hours under nitrogen. The reaction mixture is diluted with 40 ml. of ethyl acetate and filtered. The filtrate is washed sequentially with two 20 m1. portions of ice cold 10% aqueous sodium bisulfite solution, three 20 ml. portions of ice cold 5% aqueous potassium hydroxide solution and finally washed to neutrality with several 20 ml. portions of ice water. The aqueous washes are re-extracted with 50 ml. of ethyl acetate and treated as above. The combined organic phase is dried over sodium sulfate and concentrated in vacuo. The crude material dissolved in benzene is chromatographed on 15 g. of acid-washed alumina. Elution with chloroformzether 2:8 affords 11fi,17a,21-trihydroxy- 6,16a-4,6-pregnadiene3,20-dione ZI-acetate. This product is then hydrolyzed by treatment with a solution of potassium bicarbonate in aqueous methanol to form 11,8,l7a,21-trihydroxy-6,16a-dimethy1 4,6 pregnadiene- 3,20-dione.

In accordance with the above procedure but starting with 11;9,l7a,21 trihydroxy-6a,16fi-dimethyl-4-pregnene- 3,20-dione ZI-acetate there are obtained 11fi,17 x,21-trihydroxy-6,l6fldimethyl-4,6-pregnadiene-3,20-dione and its Example 28 A suspension containing 300 mg. of t-fl110IO-11f3,17a, 21-trihydroxy-6a, l 6a-dimethyl-4-pregnene-3,20-dione 21- acetate, 700 mg. of chloranil, 8.3 ml. of ethyl acetate and 1.7 ml. of acetic acid is refluxed for 17 hours under nitrogen. The reaction mixture is diluted with 40 m1. of ethyl acetate and filtered. The filtrate is washed sequentially with two 20 ml. portions of ice cold 10% aqueous sodium bisulfite solution, three 20 ml. portions of ice cold 5% aqueous potassium hydroxide solution and finally washed to neutrality with several 20 ml. portions of ice water. The aqueous washes are re-extracted with 50 ml. of ethyl acetate and treated as above. The combined organic phase is dried over sodium sulfate and concentrated in vacuo. The crude material dissolved in benzene is chromatographed on 15 g. of acid-washed alumina. Elution with chloroformzether 22'8 afiords 9u-flll0lOl1fl,l7ot,21- trihydroxy-6,16a-dimethyl-4,6-pregnadiene-3,ZO-dione 21- This'productis then hydrolyzed by treatment 

1. RING A UNSATURATED 17A-HYROXY-11-OXYGENATED-20KETO-STEROIDS OF THE PREGNANE SERIES HAVING METHYL SUBSTITUENTS ATTACHED TO BOTH THE C-6 AND C-16 CARBON ATOMS. 